Proteolysis Targeting Chimera Compounds and Methods of Preparing and Using Same

ABSTRACT

The present invention includes novel compounds and methods for preventing or treating diseases associated with and/or caused by overexpression and/or uncontrolled activation of a tyrosine kinase in a subject in need thereof. In certain embodiments, the compounds of the present invention comprise a tyrosine kinase inhibitor, a linker and a ubiquitin ligase binder. The methods of the present invention comprise administering to the subject an pharmaceutically effective amount of at least one compound of the invention.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of, and claims priority to,U.S. patent application Ser. No. 15/909,521 filed Mar. 1, 2018, which isa divisional of, and claims priority to, U.S. patent application Ser.No. 15/341,275, filed Nov. 2, 2016, now issued as U.S. Pat. No.9,938,264, which claims priority under 35 U.S.C. § 119(e) to U.S.Provisional Application No. 62/249,501, filed Nov. 2, 2015, whichapplications are hereby incorporated by reference in their entireties.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under CA197589 andAI084140 awarded by National Institutes of Health. The government hascertain rights in the invention.

BACKGROUND OF THE INVENTION

The current inhibitor-based drug paradigm not only limits drug targetsto those proteins with a tractable active site, but also requires highdosing in order to achieve adequate IC₉ concentrations for therapeuticefficacy. To circumvent these issues, alternative therapeutic strategieshave been employed to specifically knock down target proteins. Whilegenetic techniques such as RNAi, and CRISPR/Cas9 can significantlyreduce protein levels, the pharmacokinetic properties (i.e., metabolicstability and tissue distribution) associated with these approaches haveso far limited their development as clinical agents.

The pathologic fusion protein BCR-ABL is a constitutively activetyrosine kinase that drives uncontrolled cell proliferation, resultingin chronic myelogenous leukemia (CML). With the advent of tyrosinekinase inhibitors (TKIs) targeting BCR-ABL, CML has become a chronic butmanageable disease. For example, imatinib mesylate, the first TKIdeveloped against BCR-ABL, binds competitively at the ATP-binding siteof c-ABL and inhibits both c-ABL and the oncogenic fusion proteinBCR-ABL. Second generation TKIs (such as dasatinib and bosutinib) weresubsequently developed to treat CML patients with acquired resistance toimatinib. Despite the remarkable success of BCR-ABL TKIs, all CMLpatients must remain on treatment for life because the TKIs are notcurative due to persistent leukemic stem cells (LSCs).

There is thus an unmet need in the art for novel compositions andmethods to knock down c-ABL and/or BCR-ABL in a cell. Such methods couldbe used to treat and/or prevent CML in a mammal. The present inventionaddresses this need.

BRIEF SUMMARY OF THE INVENTION

The invention provides a compound of formula (I). The invention furtherprovides a pharmaceutical composition comprising at least one compoundof formula (I) and at least one pharmaceutically acceptable carrier. Theinvention further provides a method of treating or preventing a diseaseor disorder associated with overexpression and/or uncontrolledactivation of c-Abl and/or BCR-ABL. The invention further provides amethod of preventing or treating a tyrosine kinase-dependent cancer in asubject in need thereof.

In certain embodiments, the compound of formula (I) is TKI-L-(ULM)_(k)(I), wherein: TKI is a tyrosine kinase inhibitor, L is a linker, eachULM is independently a ubiquitin ligase binder, and k is an integerranging from 1 to 4, wherein TKI is covalently linked to L and whereineach ULM is covalently linked to L; or a salt, enantiomer, stereoisomer,solvate, polymorph or N-oxide thereof.

In certain embodiments, TKI is capable of binding to c-ABL and/orBCR-ABL. In other embodiments, upon binding of the compoundsimultaneously to a tyrosine kinase and a ubiquitin ligase, the tyrosinekinase is ubiquitinated by the ubiquitin ligase. In yet otherembodiments, at least one ULM binds to an E3 ubiquitin ligase. In yetother embodiments, the E3 ubiquitin ligase comprises a Von Hippel Lindau(VHL) E3 ubiquitin ligase or a Cereblon (CRBN) E3 ligase.

In certain embodiments, the TKI binds to and inhibits c-ABL. In otherembodiments, the TKI binds to and inhibits BCR-ABL. In yet otherembodiments, the TKI binds to and inhibits both c-ABL and BCR-ABL. Inyet other embodiments, the TKI is at least one selected from the groupconsisting of Dasatinib, Imatinib, Saracatinib, Ponatinib, Nilotinib,Danusertib, AT9283, Degrasyn, Bafetinib, KW-2449, NVP-BHG712, DCC-2036,GZD824, GNF-2, PD173955, GNF-5, Bosutinib, Gefitinib, Erlotinib,Sunitinib, Ruxolitinib, Tofacitinib, Lapatinib, Vandetanib, Sorafenib,Sunitinib, Axitinib, Nintedanib, Regorafenib, Pazopanib, Lenvatinib,Crizotinib, Ceritinib, Cabozantinib, DWF, Afatinib, Ibrutinib, B43,KU004, Foretinib, KRCA-0008, PF-06439015, PF-06463922, Canertinib,GSA-10, GW2974, GW583340, WZ4002, CP-380736, D2667, Mubritinib,PD153035, PD168393, Pelitinib, PF-06459988, PF-06672131, PF-6422899,PKI-166, Reveromycin A, Tyrphostin 1, Tyrphostin 23, Tyrphostin 51,Tyrphostin AG 528, Tyrphostin AG 658, Tyrphostin AG 825, Tyrphostin AG835, Tyrphostin AG 1478, Tyrphostin RG 13022, Tyrphostin RG 14620, B178,GSK1838705A, PD-161570, PD 173074, SU-5402, Roslin 2,Picropodophyllotoxin, PQ401, I-OMe-Tyrphostin AG 538, GNF 5837,GW441756, Tyrphostin AG 879, DMPQ, JNJ-10198409, PLX647, Trapidil,Tyrphostin A9, Tyrphostin AG 370, Lestaurtinib, DMH4, Geldanamycin,Genistein, GW2580, Herbimycin A, Lavendustin C, Midostaurin, NVP-BHG712,PD158780, PD-166866, PF-06273340, PP2, RPI, SU 11274, SU5614, Symadex,Tyrphostin AG 34, Tyrphostin AG 974, Tyrphostin AG 1007, UNC2881,Honokiol, SU1498, SKLB1002, CP-547632, JK-P3, KRN633, SC-1, ST638, SU5416, Sulochrin, Tyrphostin SU 1498, S8567, rociletinib, Dacomitinib,Tivantinib, Neratinib, Masitinib, Vatalanib, Icotinib, XL-184, OSI-930,AB1010, Quizartinib, AZD9291, Tandutinib, HM61713, Brigantinib,Vemurafenib (PLX-4032), Semaxanib, AZD2171, Crenolanib, Damnacanthal,Fostamatinib, Motesanib, Radotinib, OSI-027, Linsitinib, BIX02189,PF-431396, PND-1186, PF-03814735, PF-431396, sirolimus, temsirolimus,everolimus, deforolimus, zotarolimus, BEZ235, INK128, Omipalisib,AZD8055, MHY1485, PI-103, KU-0063794, ETP-46464, GDC-0349, XL388,WYE-354, WYE-132, GSK1059615, WAY-600, PF-04691502, WYE-687, PP121,BGT226, AZD2014, PP242, CH5132799, P529, GDC-0980, GDC-0994, XMD8-92,Ulixertinib, FR180204, SCH772984, Trametinib, PD184352, PD98059,Selumetinib, PD325901, U0126, Pimasertinib, TAK-733, AZD8330,Binimetinib, PD318088, SL-327, Refametinib, GDC-0623, Cobimetinib,BI-847325, Adaphostin, GNF 2, PPY A, AIM-100, ASP 3026, LFM A13, PF06465469, (−)-Terreic acid, AG-490, BIBU 1361, BIBX 1382, BMS 599626,CGP 52411, GW 583340, HDS 029, HKI 357, JNJ 28871063, WHI-P 154, PF431396, PF 573228, FIN 1, PD 166285, SUN 11602, SR 140333, TCS 359, BMS536924, NVP ADW 742, PQ 401, BMS 509744, CP 690550, NSC 33994, WHI-P154, KB SRC 4, DDR1-IN-1, PF 04217903, PHA 665752, SU 16f, A 419259, AZM475271, PP 1, PP 2, 1-Naphthyl PP1, Src I1, ANA 12, PD 90780, Ki 8751,Ki 20227, ZM 306416, ZM 323881, AEE 788, GTP 14564, PD 180970, R 1530,SU 6668, Toceranib, CEP-32496 (1-(3-((6,7-dimethoxyquinazolin-4-yl)oxy)phenyl)-3-(5-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazol-3-yl)urea),AZ 628 (4-(2-cyanopropan-2-yl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)benzamide), Vemurafenib (PLX-4032), PLX-4720(N-(3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide),SB 590885 ((E)-5-(2-(4-(2-(dimethylamino)ethoxy)phenyl)-4-(pyridin-4-yl)-1H-imidazol-5-yl)-2,3-dihydro-1H-inden-1-oneoxime), GDC-0879((E)-5-(2-(2-hydroxyethyl)-4-(pyridin-4-yl)-1H-imidazol-5-yl)-2,3-dihydro-1H-inden-1-oneoxime), a compound of formula

wherein R¹ is H or CH₃, and R² is

a compound of formula

wherein R is

and a compound of formula

wherein the broken lines correspond to the divalent group

In yet other embodiments, the TKI is Imatinib, Dasatinib or Bosutinib.

In certain embodiments, at least one ULM comprises formula (IX):

In other embodiments, at least one ULM comprises formula (X):

In yet other embodiments, k=1.

In certain embodiments, the linker L corresponds to formula—(CH₂)_(m1)—X₄—(CH₂—CH₂—X₅)_(m2)—(CH₂)_(m3)—C(X₆)—, wherein: —(CH₂)_(m1)is covalently bound to the TKI, and C(X₃)— is covalently bound to theULM; wherein each m1, m2, and m3 is independently 0, 1, 2, 3, 4, 5, 6,7, 8, 9, or 10; wherein each X₄, X₅, and X₆ is independently absent (abond), O, S, or N—R²⁰, wherein each R²⁰ is independently selected fromthe group consisting of hydrogen, optionally substituted C₁-C₆ alkyl,optionally substituted aryl, optionally substituted heteroaryl,optionally substituted C₃-C₈ cycloalkyl, and optionally substitutedC₃-C₈ cycloheteroalky. In other embodiments, m is 6; m2 is 1 or 2; m3 is1 or 5; and X₄, X₅, and X₆ are O. In yet other embodiments, m is 6; m2is 5; m3 is 5; X₄ and X₆ are 0; and X₅ is absent. In yet otherembodiments, m is 6; m2 is 5; m3 is 1; X₄, X₅, and X₆ are O.

In certain embodiments, the compound is selected from the groupconsisting of:N-(2-chloro-6-methylphenyl)-2-((6-(4-((S)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-11,14,17-trioxa-4-azatricosan-23-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-6-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-22-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-2-2-6-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-VHL):

(2S,4R)-1-((S)-3,3-dimethyl-2-(2-(2-((6-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-2-2-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-((S)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19,22-hexaoxa-4-azaoctacosan-28-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-2-2-2-2-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-27-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-2-2-2-2-2-2-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-((5-((6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-5-6-VHL):

(2S,4R)-1-((S)-3,3-dimethyl-2-(6-((5-((6-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-5-6-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-22-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-2-2-6-VHL):

(2S,4R)-1-((S)-2-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-2-2-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-27-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-2-2-2-2-2-2-VHL):

(2S,4R)-1-((S)-2-(6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-5-6-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)ethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-6-CRBN):

6-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide(BOS-6-2-2-6-CRBN):

4-((4-(6-(2-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)ethoxy)ethoxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide(IMA-6-2-2-6-CRBN):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-CRBN):

2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide(BOS-6-2-2-CRBN):

4-((4-(6-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide(IMA-6-2-2-CRBN):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-1-oxo-3,6,9,12,15,18-hexaoxatetracosan-24-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-2-2-2-2-CRBN):

24-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-3,6,9,12,15,18-hexaoxatetracosanamide(BOS-6-2-2-2-2-2-2-CRBN):

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-24-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxatetracosanamide(IMA-6-2-2-2-2-2-2-CRBN):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide_(DAS-6-5-6-CRBN):

6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide(BOS-6-5-6-CRBN):

4-((4-(6-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide(MA-6-5-6-CRBN):

In certain embodiments, the pharmaceutical composition further comprisesat least one additional therapeutic compound that treats or preventscancer.

In certain embodiments, the method comprises administering to thesubject a therapeutically effective amount of at least one compound ofthe invention. In other embodiments, the disease or disorder comprisescancer. In yet other embodiments, the cancer comprises chronicmyelogenous leukemia (CML). In yet other embodiments, the compound isadministered to the subject by at least one route selected from thegroup consisting of nasal, inhalational, topical, oral, buccal, rectal,pleural, peritoneal, vaginal, intramuscular, subcutaneous, transdermal,epidural, intrathecal and intravenous routes. In yet other embodiments,the cancer is associated with overexpression and/or uncontrolledactivation of the tyrosine kinase. In other embodiments, the tyrosinekinase is oncogenic. In yet other embodiments, the subject is a human.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of specific embodiments of theinvention will be better understood when read in conjunction with theappended drawings. For the purpose of illustrating the invention, thereare shown in the drawings specific embodiments. It should be understood,however, that the invention is not limited to the precise arrangementsand instrumentalities of the embodiments shown in the drawings.

FIGS. 1A-1C comprise illustrations of a non-limiting approach toproteolysis targeting chimera (PROTAC) development. FIG. 1A comprises animage illustrating that PROTACs act through proximity-inducedubiquitination, leading to subsequent degradation by the proteasome.FIG. 1B comprises an image illustrating overlay of the TKIs bosutinib(blue; PDB: 3UE4) and dasatinib (yellow; PDB: 2GQG), in the ABLATP-binding pocket. Linkers are attached via the solvent exposed site(red circle). FIG. 1C comprises linkers utilized to connect therespective TKI to the E3 recruiting ligand.

FIGS. 2A-2B illustrate biological effects of VHL-based PROTACs.BOS-6-2-2-6-VHL (FIG. 2A) and DAS-6-2-2-6-VHL (FIG. 2B) were incubatedwith K562 human chronic myelogenous leukemia cells for 24 hrs. Theconcentrations of the parent inhibitors were 1 μM. As determined byimmunoblot, degradation of c-ABL was observed with DAS-VHL starting at 1μM; however, no degradation of BCR-ABL was observed in any of theVHL-based PROTACs.

FIGS. 3A-3B illustrate biological effects of CRBN-based PROTACs.BOS-6-2-2-6-CRBN (FIG. 3A) and DAS-6-2-2-6-CRBN (FIG. 3B) were incubatedwith K562 cells for 24 hrs. The concentrations of the parent inhibitorswere 1 μM. As determined by immunoblot, degradation of BCR-ABL and c-ABLwas observed in the DAS-CRBN and BOS-CRBN series.

FIG. 4 comprises a graph illustrating cell viability withDAS-6-2-2-6-CRBN. This PROTAC was greater than a thousand-fold moreeffective against the BCR-ABL driven cell line K562 over non-BCR-ABLdriven cell lines, as determined by CellTiter-Glo® Luminescent CellViability assay after a 48 hr treatment. Error bars displayed are S.E.M(n=3). Data was normalized to DMSO-treated controls.

FIG. 5 comprises a schematic illustration of the finding thatcombination of the inhibitor warhead and the recruited E3 ubiquitinligase permits targets to be accessed for degradation. IMA-based PROTACsdid not induce the degradation of c-ABL or BCR-ABL, despite targetengagement.

FIGS. 6A-6B illustrate IMA-based PROTACs with linker 6-2-2-6.IMA-6-2-2-6-VHL (FIG. 6A) and IMA-6-2-2-6-CRBN (FIG. 6B) were incubatedwith K562 human chronic myelogenous leukemia cell line for 24 hrs. Theconcentration of the parent inhibitors were at 1 μM. As determined byimmunoblot, no degradation of c-ABL or BCR-ABL was observed in any ofthe IMA-based PROTACs.

FIGS. 7A-7F illustrate PROTACs with linker 6-5-6. IMA-6-5-6-VHL (FIG.7A), BOS-6-5-6-VHL (FIG. 7B) and DAS-6-5-6-VHL (FIG. 7C) were incubatedwith K562 human chronic myelogenous leukemia cell line for 24 hrs.IMA-6-5-6-CRBN (FIG. 7D), BOS-6-5-6-CRBN (FIG. 7E) and DAS-6-5-6-CRBN(FIG. 7F) were incubated with K562 human chronic myelogenous leukemiacell line for 24 hrs. The concentration of the parent inhibitors were at1 μM. As determined by immunoblot, degradation of c-ABL can be observedwith DAS-6-5-6-VHL starting at 1 μM. However, no degradation of BCR-ABLwas observed in any of the VHL-based PROTACs.

FIGS. 8A-8F illustrates PROTACs with linker 6-2-2. IMA-6-2-2-VHL (FIG.8A), BOS-6-2-2-VHL (FIG. 8B) and DAS-6-2-2-VHL (FIG. 8C) were incubatedwith K562 human chronic myelogenous leukemia cell line for 24 hrs.IMA-6-2-2-CRBN (FIG. 8D), BOS-6-2-2-CRBN (FIG. 8E) and DAS-6-2-2-CRBN(FIG. 8F) were incubated with K562 human chronic myelogenous leukemiacell line for 24 hrs. The concentration of the parent inhibitors were at1 μM. As determined by immunoblot, degradation of c-ABL can be observedwith DAS-6-2-2-VHL starting at 1 μM. However, no degradation of BCR-ABLwas observed in any of the VHL-based PROTACs.

FIGS. 9A-9F illustrates PROTACs with linker 6-(2)₅-2. IMA-6-(2)₅-2-VHL(FIG. 9A), BOS-6-(2)₅-2-VHL (FIG. 9B) and DAS-6-(2)₅-2-VHL (FIG. 9C)were incubated with K562 human chronic myelogenous leukemia cell linefor 24 hrs. IMA-6-(2)₅-2-CRBN (FIG. 9D), BOS-6-(2)₅-2-CRBN (FIG. 9E) andDAS-6-(2)₅-2-CRBN (FIG. 9F) were incubated with K562 human chronicmyelogenous leukemia cell line for 24 hrs. The concentration of theparent inhibitors were at 1 μM. As determined by immunoblot, degradationof c-ABL can be observed with DAS-6-(2)₅-2-VHL starting at 1 μM.However, no degradation of BCR-ABL was observed in any of the VHL-basedPROTACs.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the unexpected discovery ofbifunctional small compounds that efficiently degrade certaincancer-related tyrosine kinases in a cellular environment. Thesecompounds are based on proteolysis targeting chimera (PROTAC)technology, wherein one end of the compound recruits a ubiquitin ligasewhile the other end engages the target tyrosine kinase. In certainembodiments, the ubiquitin ligase is an E3 ubiquitin ligase. In otherembodiments, the ubiquitin ligase is Von Hippel Lindau (VHL) E3ubiquitin ligase and/or Cereblon (CRBN) E3 ligase. Ternary complexformation takes place when the compounds of the invention bind to thetyrosine kinase and the ubiquitin ligase, thus bringing the recruitedligase in close proximity with the tyrosine kinase. This leads to theubiquitination of the tyrosine kinase of interest and its subsequentdegradation by proteasome.

In certain embodiments, the compounds of the invention can be used totreat diseases associated with overexpression and/or uncontrolledactivation of the tyrosine kinase. In other embodiments, the compoundsof the invention can be used to treat a cancer that is associated withand/or caused by an oncogenic tyrosine kinase.

As demonstrated herein, bifunctional small compounds based on two potentTKIs (bosutinib and dasatinib) were synthesized and shown to mediate thedegradation of c-ABL and BCR-ABL by hijacking either CRBN or VHL E3ubiquitin ligase. Furthermore, these novel PROTACs were shown to beselective against the BCR-ABL driven cell line K562. In certainembodiments, changing the inhibitor warhead and the recruited E3 ligaseinfluences which protein targets are susceptible to PROTAC-induceddegradation (FIG. 5). By varying the recruited E3 ligase, the substratespectrum of PROTACs can be significantly altered. In certainembodiments, the selectivity of a promiscuous inhibitor may be narrowedby creating a more selective degrader via the coupling to different E3ligase recruiting ligands.

The present description provides compounds comprising a ligand, e.g., asmall molecule ligand (i.e., having a molecular weight that is lowerthan about 2,000, 1,000, 500, or 200 Daltons), which is capable ofbinding to a ubiquitin ligase, such as, but not limited to, VHL orcereblon. The compounds also comprise a moiety that is capable ofbinding to a target protein, in such a way that the target protein isplaced in proximity to the ubiquitin ligase to effect degradation(and/or inhibition) of that protein. In certain embodiments, “smallmolecule” means, in addition to the above, that the molecule isnon-peptidyl, i.e., it is not generally considered a peptide, e.g.,comprises fewer than 4, 3, or 2 amino acids. In accordance with thepresent description, the PTM, ULM and/or PROTAC molecules can be a smallmolecule.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, specific methods andmaterials are described.

As used herein, each of the following terms has the meaning associatedwith it in this section.

The articles “a” and “an” are used herein to refer to one or to morethan one (i.e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

“About” as used herein when referring to a measurable value such as anamount, a temporal duration, and the like, is meant to encompassvariations of 20% or 10%, more preferably 5%, even more preferably +1%,and still more preferably 0.1% from the specified value, as suchvariations are appropriate to perform the disclosed methods.

The term “abnormal” when used in the context of organisms, tissues,cells or components thereof, refers to those organisms, tissues, cellsor components thereof that differ in at least one observable ordetectable characteristic (e.g., age, treatment, time of day, etc.) fromthose organisms, tissues, cells or components thereof that display the“normal” (expected) respective characteristic. Characteristics that arenormal or expected for one cell or tissue type might be abnormal for adifferent cell or tissue type.

A disease or disorder is “alleviated” if the severity of a symptom ofthe disease or disorder, the frequency with which such a symptom isexperienced by a patient, or both, is reduced.

The terms “cancer” refers to the physiological condition in a subjecttypically characterized by unregulated cell growth. Examples of cancerinclude, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma,and leukemia or lymphoid malignancies. More particular examples of suchcancers include squamous cell cancer (e.g., epithelial squamous cellcancer), lung cancer including small cell lung cancer, non-small celllung cancer (“NSCLC”), vulval cancer, thyroid cancer, adenocarcinoma ofthe lung and squamous carcinoma of the lung, cancer of the peritoneum,hepatocellular cancer, gastric or stomach cancer includinggastrointestinal cancer, pancreatic cancer, glioblastoma, cervicalcancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breastcancer, colon cancer, rectal cancer, colorectal cancer, endometrial oruterine carcinoma, salivary gland carcinoma, kidney or renal cancer,prostate cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, aswell as head and neck cancer. In yet other embodiments, the cancer is atleast one selected from the group consisting of ALL, T-lineage Acutelymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL),Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-BLymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL,Philadelphia chromosome positive ALL, Philadelphia chromosome positiveCML, lymphoma, leukemia, multiple myeloma myeloproliferative diseases,large B cell lymphoma, and B cell Lymphoma. Without wishing to belimited by any theory, in about 10% of patients with acute lymphocyticleukemia, patients carry a 9;22 translocation cytogeneticallyindistinguishable from the Philadelphia chromosome of CML.

As used herein, the term “composition” or “pharmaceutical composition”refers to a mixture of at least one compound useful within the inventionwith a pharmaceutically acceptable carrier. The pharmaceuticalcomposition facilitates administration of the compound to a patient orsubject. Multiple techniques of administering a compound exist in theart including, but not limited to, intravenous, oral, aerosol,parenteral, ophthalmic, pulmonary and topical administration.

A “disease” is a state of health of an animal wherein the animal cannotmaintain homeostasis, and wherein if the disease is not ameliorated thenthe animal's health continues to deteriorate.

In contrast, a “disorder” in an animal is a state of health in which theanimal is able to maintain homeostasis, but in which the animal's stateof health is less favorable than it would be in the absence of thedisorder. Left untreated, a disorder does not necessarily cause afurther decrease in the animal's state of health.

As used herein, the terms “effective amount,” “pharmaceuticallyeffective amount” and “therapeutically effective amount” refer to anontoxic but sufficient amount of an agent to provide the desiredbiological result. That result may be reduction and/or alleviation ofthe signs, symptoms, or causes of a disease, or any other desiredalteration of a biological system. An appropriate therapeutic amount inany individual case may be determined by one of ordinary skill in theart using routine experimentation.

As used herein, the term “efficacy” refers to the maximal effect (E_(m))achieved within an assay.

As used herein, the term “L” or “Linker” refers to the linker.

As used herein, the term “pharmaceutically acceptable” refers to amaterial, such as a carrier or diluent, which does not abrogate thebiological activity or properties of the compound, and is relativelynon-toxic, i.e., the material may be administered to an individualwithout causing undesirable biological effects or interacting in adeleterious manner with any of the components of the composition inwhich it is contained.

As used herein, the language “pharmaceutically acceptable salt” refersto a salt of the administered compounds prepared from pharmaceuticallyacceptable non-toxic acids or bases, including inorganic acids or bases,organic acids or bases, solvates, hydrates, or clathrates thereof.

Suitable pharmaceutically acceptable acid addition salts may be preparedfrom an inorganic acid or from an organic acid. Examples of inorganicacids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic,sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids(including hydrogen phosphate and dihydrogen phosphate). Appropriateorganic acids may be selected from aliphatic, cycloaliphatic, aromatic,araliphatic, heterocyclic, carboxylic and sulfonic classes of organicacids, examples of which include formic, acetic, propionic, succinic,glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic,glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic,mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic,benzenesulfonic, pantothenic, trifluoromethanesulfonic,2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic,galactaric and galacturonic acid.

Suitable pharmaceutically acceptable base addition salts of compounds ofthe invention include, for example, ammonium salts, metallic saltsincluding alkali metal, alkaline earth metal and transition metal saltssuch as, for example, calcium, magnesium, potassium, sodium and zincsalts. Pharmaceutically acceptable base addition salts also includeorganic salts made from basic amines such as, for example,N,N′-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (N-methylglucamine) and procaine. All ofthese salts may be prepared from the corresponding compound by reacting,for example, the appropriate acid or base with the compound.

As used herein, the term “pharmaceutically acceptable carrier” means apharmaceutically acceptable material, composition or carrier, such as aliquid or solid filler, stabilizer, dispersing agent, suspending agent,diluent, excipient, thickening agent, solvent or encapsulating material,involved in carrying or transporting a compound useful within theinvention within or to the patient such that it may perform its intendedfunction. Typically, such constructs are carried or transported from oneorgan, or portion of the body, to another organ, or portion of the body.Each carrier must be “acceptable” in the sense of being compatible withthe other ingredients of the formulation, including the compound usefulwithin the invention, and not injurious to the patient. Some examples ofmaterials that may serve as pharmaceutically acceptable carriersinclude: sugars, such as lactose, glucose and sucrose; starches, such ascorn starch and potato starch; cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients, such as cocoabutter and suppository waxes; oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols,such as propylene glycol; polyols, such as glycerin, sorbitol, mannitoland polyethylene glycol; esters, such as ethyl oleate and ethyl laurate;agar; buffering agents, such as magnesium hydroxide and aluminumhydroxide; surface active agents; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffersolutions; and other non-toxic compatible substances employed inpharmaceutical formulations. As used herein, “pharmaceuticallyacceptable carrier” also includes any and all coatings, antibacterialand antifungal agents, and absorption delaying agents, and the like thatare compatible with the activity of the compound useful within theinvention, and are physiologically acceptable to the patient.Supplementary active compounds may also be incorporated into thecompositions. The “pharmaceutically acceptable carrier” may furtherinclude a pharmaceutically acceptable salt of the compound useful withinthe invention. Other additional ingredients that may be included in thepharmaceutical compositions used in the practice of the invention areknown in the art and described, for example in Remington'sPharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton,Pa.), which is incorporated herein by reference.

The terms “patient,” “subject,” or “individual” are used interchangeablyherein, and refer to any animal, or cells thereof whether in vitro or insitu, amenable to the methods described herein. In a non-limitingembodiment, the patient, subject or individual is a human.

As used herein, the term “potency” refers to the dose needed to producehalf the maximal response (ED₅₀).

A “therapeutic” treatment is a treatment administered to a subject whoexhibits signs of pathology, for the purpose of diminishing oreliminating those signs.

As used herein, the term “treatment” or “treating” is defined as theapplication or administration of a therapeutic agent, i.e., a compoundof the invention (alone or in combination with another pharmaceuticalagent), to a patient, or application or administration of a therapeuticagent to an isolated tissue or cell line from a patient (e.g., fordiagnosis or ex vivo applications), who has a condition contemplatedherein, a symptom of a condition contemplated herein or the potential todevelop a condition contemplated herein, with the purpose to cure, heal,alleviate, relieve, alter, remedy, ameliorate, improve or affect acondition contemplated herein, the symptoms of a condition contemplatedherein or the potential to develop a condition contemplated herein. Suchtreatments may be specifically tailored or modified, based on knowledgeobtained from the field of pharmacogenomics.

As used herein, the term “alkyl,” by itself or as part of anothersubstituent means, unless otherwise stated, a straight or branched chainhydrocarbon having the number of carbon atoms designated (i.e. C₁-6means one to six carbon atoms) and including straight, branched chain,or cyclic substituent groups. Examples include methyl, ethyl, propyl,isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, andcyclopropylmethyl. Most preferred is (C₁-C₆)alkyl, particularly ethyl,methyl, isopropyl, isobutyl, n-pentyl, n-hexyl and cyclopropylmethyl.

As used herein, the term “substituted alkyl” means alkyl as definedabove, substituted by one, two or three substituents selected from thegroup consisting of halogen, —OH, alkoxy, —NH₂, —N(CH₃)₂, —C(═O)OH,trifluoromethyl, —C≡N, —C(═O)O(C₁-C₄)alkyl, —C(═O)NH₂, —SO₂NH₂,—C(═NH)NH₂, and —NO₂, preferably containing one or two substituentsselected from halogen, —OH, alkoxy, —NH₂, trifluoromethyl, —N(CH₃)₂, and—C(═O)OH, more preferably selected from halogen, alkoxy and —OH.Examples of substituted alkyls include, but are not limited to,2,2-difluoropropyl, 2-carboxycyclopentyl and 3-chloropropyl.

As used herein, the term “haloalkyl” means alkyl as defined above,substituted by one, two or three substituents selected from the groupconsisting of F, Cl, Br, and I.

As used herein, the term “heteroalkyl” by itself or in combination withanother term means, unless otherwise stated, a stable straight orbranched chain alkyl group consisting of the stated number of carbonatoms and one or two heteroatoms selected from the group consisting ofO, N, and S, and wherein the nitrogen and sulfur atoms may be optionallyoxidized and the nitrogen heteroatom may be optionally quaternized orsubstituted. The heteroatom(s) may be placed at any position of theheteroalkyl group, including between the rest of the heteroalkyl groupand the fragment to which it is attached, as well as attached to themost distal carbon atom in the heteroalkyl group. Examples include:—CH₂—CH₂—CH₃, —CH₂—CH₂—CH₂—OH, —CH₂—CH₂—NH—CH₃, —CH₂—S—CH₂—CH₃,—NH—(CH₂)_(m)OH (m=1-6), —N(CH₃)—(CH₂)_(m)—OH (m=1-6),—NH—(CH₂)_(m)—OCH₃ (m=1-6), and —CH₂CH₂—S(═O)—CH₃. Up to two heteroatomsmay be consecutive, such as, for example, —CH₂—NH—OCH₃, or—CH₂—CH₂—S—S—CH₃

As used herein, the term “alkoxy” employed alone or in combination withother terms means, unless otherwise stated, an alkyl group having thedesignated number of carbon atoms, as defined above, connected to therest of the molecule via an oxygen atom, such as, for example, methoxy,ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs andisomers. Preferred are (C₁-C₃) alkoxy, particularly ethoxy and methoxy.

As used herein, the term “cycloalkyl” refers to a mono cyclic orpolycyclic non-aromatic radical, wherein each of the atoms forming thering (i.e. skeletal atoms) is a carbon atom. In certain embodiments, thecycloalkyl group is saturated or partially unsaturated. In otherembodiments, the cycloalkyl group is fused with an aromatic ring.Cycloalkyl groups include groups having from 3 to 10 ring atoms.Illustrative examples of cycloalkyl groups include, but are not limitedto, the following moieties:

Monocyclic cycloalkyls include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.Dicyclic cycloalkyls include, but are not limited to,tetrahydronaphthyl, indanyl, and tetrahydropentalene. Polycycliccycloalkyls include adamantine and norbornane. The term cycloalkylincludes “unsaturated nonaromatic carbocyclyl” or “nonaromaticunsaturated carbocyclyl” groups, both of which refer to a nonaromaticcarbocycle as defined herein, which contains at least one carbon carbondouble bond or one carbon carbon triple bond.

As used herein, the term “aromatic” refers to a carbocycle orheterocycle with one or more polyunsaturated rings and having aromaticcharacter, i.e. having (4n+2) delocalized 7 (pi) electrons, where n isan integer.

As used herein, the term “aryl,” employed alone or in combination withother terms, means, unless otherwise stated, a carbocyclic aromaticsystem containing one or more rings (typically one, two or three rings),wherein such rings may be attached together in a pendent manner, such asa biphenyl, or may be fused, such as naphthalene. Examples of arylgroups include phenyl, anthracyl, and naphthyl. Preferred examples arephenyl and naphthyl, most preferred is phenyl.

As used herein, the term “aryl-(C₁-C₃)alkyl” means a functional groupwherein a one- to three-carbon alkylene chain is attached to an arylgroup, e.g., —CH₂CH₂-phenyl. Preferred is aryl-CH₂— and aryl-CH(CH₃)—.The term “substituted aryl-(C₁-C₃)alkyl” means an aryl-(C₁-C₃)alkylfunctional group in which the aryl group is substituted. Preferred issubstituted aryl(CH₂)—. Similarly, the term “heteroaryl-(C₁-C₃)alkyl”means a functional group wherein a one to three carbon alkylene chain isattached to a heteroaryl group, e.g., —CH₂CH₂-pyridyl. Preferred isheteroaryl-(CH₂)—. The term “substituted heteroaryl-(C₁-C₃)alkyl” meansa heteroaryl-(C₁-C₃)alkyl functional group in which the heteroaryl groupis substituted. Preferred is substituted heteroaryl-(CH₂)—.

As used herein, the term “halo” or “halogen” alone or as part of anothersubstituent means, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom, preferably, fluorine, chlorine, or bromine,more preferably, fluorine or chlorine.

As used herein, the term “heterocycloalkyl” or “heterocyclyl” refers toa heteroalicyclic group containing one to four ring heteroatoms eachselected from O, S and N. In certain embodiments, each heterocycloalkylgroup has from 4 to 10 atoms in its ring system, with the proviso thatthe ring of said group does not contain two adjacent O or S atoms. Inother embodiments, the heterocycloalkyl group is fused with an aromaticring. In certain embodiments, the nitrogen and sulfur heteroatoms may beoptionally oxidized, and the nitrogen atom may be optionallyquaternized. The heterocyclic system may be attached, unless otherwisestated, at any heteroatom or carbon atom that affords a stablestructure. A heterocycle may be aromatic or non-aromatic in nature. Incertain embodiments, the heterocycle is a heteroaryl.

An example of a 3-membered heterocycloalkyl group includes, and is notlimited to, aziridine. Examples of 4-membered heterocycloalkyl groupsinclude, and are not limited to, azetidine and a beta lactam. Examplesof 5-membered heterocycloalkyl groups include, and are not limited to,pyrrolidine, oxazolidine and thiazolidinedione. Examples of 6-memberedheterocycloalkyl groups include, and are not limited to, piperidine,morpholine and piperazine. Other non-limiting examples ofheterocycloalkyl groups are:

Examples of non-aromatic heterocycles include monocyclic groups such asaziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine,pyrroline, pyrazolidine, imidazoline, dioxolane, sulfolane,2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophane,piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine,morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran,1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane,4,7-dihydro-1,3-dioxepin, and hexamethyleneoxide.

As used herein, the term “heteroaryl” or “heteroaromatic” refers to aheterocycle having aromatic character. A polycyclic heteroaryl mayinclude one or more rings that are partially saturated. Examples includethe following moieties.

Examples of heteroaryl groups also include pyridyl, pyrazinyl,pyrimidinyl (particularly 2- and 4-pyrimidinyl), pyridazinyl, thienyl,furyl, pyrrolyl (particularly 2-pyrrolyl), imidazolyl, thiazolyl,oxazolyl, pyrazolyl (particularly 3- and 5-pyrazolyl), isothiazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl,1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyl and1,3,4-oxadiazolyl.

Examples of polycyclic heterocycles and heteroaryls include indolyl(particularly 3-, 4-, 5-, 6- and 7-indolyl), indolinyl, quinolyl,tetrahydroquinolyl, isoquinolyl (particularly 1- and 5-isoquinolyl),1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl (particularly 2-and 5-quinoxalinyl), quinazolinyl, phthalazinyl, 1,8-naphthyridinyl,1,4-benzodioxanyl, coumarin, dihydrocoumarin, 1,5-naphthyridinyl,benzofuryl (particularly 3-, 4-, 5-, 6- and 7-benzofuryl),2,3-dihydrobenzofuryl, 1,2-benzisoxazolyl, benzothienyl (particularly3-, 4-, 5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl(particularly 2-benzothiazolyl and 5-benzothiazolyl), purinyl,benzimidazolyl (particularly 2-benzimidazolyl), benzotriazolyl,thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrrolizidinyl, andquinolizidinyl.

As used herein, the term “substituted” means that an atom or group ofatoms has replaced hydrogen as the substituent attached to anothergroup. The term “substituted” further refers to any level ofsubstitution, namely mono-, di-, tri-, tetra-, or penta-substitution,where such substitution is permitted. The substituents are independentlyselected, and substitution may be at any chemically accessible position.In certain embodiments, the substituents vary in number between one andfour. In other embodiments, the substituents vary in number between oneand three. In yet other embodiments, the substituents vary in numberbetween one and two.

As used herein, the term “optionally substituted” means that thereferenced group may be substituted or unsubstituted. In certainembodiments, the referenced group is optionally substituted with zerosubstituents, i.e., the referenced group is unsubstituted. In otherembodiments, the referenced group is optionally substituted with one ormore additional group(s) individually and independently selected fromgroups described herein.

In certain embodiments, the substituents are independently selected fromthe group consisting of oxo, halogen, —CN, —NH₂, —OH, —NH(CH₃),—N(CH₃)₂, alkyl (including straight chain, branched and/or unsaturatedalkyl), substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, fluoro alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted alkoxy,fluoroalkoxy, —S-alkyl, S(═O)₂ alkyl, —C(═O)NH[substituted orunsubstituted alkyl, or substituted or unsubstituted phenyl], —C(═O)N[Hor alkyl]₂, —OC(═O)N[substituted or unsubstituted alkyl]₂,—NHC(═O)NH[substituted or unsubstituted alkyl, or substituted orunsubstituted phenyl], —NHC(═O)alkyl, —N[substituted or unsubstitutedalkyl]C(═O)[substituted or unsubstituted alkyl], —NHC(═O)[substituted orunsubstituted alkyl], —C(OH)[substituted or unsubstituted alkyl]₂, and—C(NH₂)[substituted or unsubstituted alkyl]₂. In other embodiments, byway of example, an optional substituent is selected from oxo, fluorine,chlorine, bromine, iodine, —CN, —NH₂, —OH, —NH(CH₃), —N(CH₃)₂, —CH₃,—CH₂CH₃, —CH(CH₃)₂, —CF₃, —CH₂CF₃, —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —OCF₃,—OCH₂CF₃, —S(═O)₂—CH₃, —C(═O)NH₂, —C(═O)—NHCH₃, —NHC(═O)NHCH₃,—C(═O)CH₃, and —C(═O)OH. In yet one embodiment, the substituents areindependently selected from the group consisting of C₁₋₆ alkyl, —OH,C₁₋₆ alkoxy, halo, amino, acetamido, oxo and nitro. In yet otherembodiments, the substituents are independently selected from the groupconsisting of C₁₋₆ alkyl, C₁₋₆ alkoxy, halo, acetamido, and nitro. Asused herein, where a substituent is an alkyl or alkoxy group, the carbonchain may be branched, straight or cyclic, with straight beingpreferred.

Ranges: throughout this disclosure, various aspects of the invention canbe presented in a range format. It should be understood that thedescription in range format is merely for convenience and brevity andshould not be construed as an inflexible limitation on the scope of theinvention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible sub-ranges as well asindividual numerical values within that range. For example, descriptionof a range such as from 1 to 6 should be considered to have specificallydisclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numberswithin that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. Thisapplies regardless of the breadth of the range.

Compounds

The compounds of the present invention may be synthesized usingtechniques well-known in the art of organic synthesis. The startingmaterials and intermediates required for the synthesis may be obtainedfrom commercial sources or synthesized according to methods known tothose skilled in the art. General procedure of making certain compoundsof the invention is described in U.S. patent application Ser. No.14/371,956, which is incorporated by reference in its entirety.

The present invention provides a compound of formula (I), or a salt,enantiomer, stereoisomer, solvate, polymorph or N-oxide thereof:TKI-L-(ULM)_(k) (I), wherein TKI is a tyrosine kinase inhibitor; L is alinker; ULM is a ubiquitin ligase binder; and k is an integer rangingfrom 1 to 4; wherein TKI is covalently linked to L and ULM is covalentlylinked to L. In certain embodiments, TKI is capable of binding thetyrosine kinase c-ABL and/or BCR-ABL, wherein, upon binding of thetyrosine kinase to the compound, the tyrosine kinase is ubiquitinated bya ubiquitin ligase.

In any of the aspects or embodiments described herein, the PTM and/orULM have an affinity (IC₅₀) for their respective target protein of lessthan about 500 μM, 450 μM, 400 μM, 350 μM, 300 μM, 250 μM, 200 μM, 150μM, 100 μM, 50 μM, 10 μM, 0.10 μM, 0.01 μM, 0.001 μM, 0.1 nM, 0.01 nM,0.001 nM or less. The determination of the IC₅₀ can be performed usingmethods well known to those of skill in the art in view of the presentdisclosure.

Tyrosine Kinase Inhibitor (TKI)

A tyrosine kinase inhibitor (TKI) contemplated within the presentinvention binds to and inhibits a tyrosine kinase, or a subunit thereof.In certain embodiments, the TKI of the present invention binds to andinhibits c-ABL. In other embodiments, the TKI of the present inventionbinds to and inhibits BCR-ABL. In yet other embodiments, the TKI of thepresent invention binds to and inhibits c-ABL and BCR-ABL.

In certain embodiments, the TKI of the invention is selected from thegroup consisting of Dasatinib, Imatinib, Saracatinib, Ponatinib,Nilotinib, Danusertib, AT9283, Degrasyn, Bafetinib, KW-2449, NVP-BHG712,DCC-2036, GZD824, GNF-2, PD173955, GNF-5, Bosutinib, Gefitinib,Erlotinib, and Sunitinib.

In certain embodiments, the TKI of the invention is selected from thegroup consisting of Ruxolitinib, Tofacitinib, Lapatinib, Vandetanib,Sorafenib, Sunitinib, Axitinib, Nintedanib, Regorafenib, Pazopanib,Lenvatinib, Crizotinib, Ceritinib, Cabozantinib, DWF, Afatinib,Ibrutinib, B43, KU004, Foretinib, KRCA-0008, PF-06439015, PF-06463922,Canertinib, GSA-10, GW2974, GW583340, WZ4002, CP-380736, D2667,Mubritinib, PD153035, PD168393, Pelitinib, PF-06459988, PF-06672131,PF-6422899, PKI-166, Reveromycin A, Tyrphostin 1, Tyrphostin 23,Tyrphostin 51, Tyrphostin AG 528, Tyrphostin AG 658, Tyrphostin AG 825,Tyrphostin AG 835, Tyrphostin AG 1478, Tyrphostin RG 13022, TyrphostinRG 14620, B178, GSK1838705A, PD-161570, PD 173074, SU-5402, Roslin 2,Picropodophyllotoxin, PQ401, I-OMe-Tyrphostin AG 538, GNF 5837,GW441756, Tyrphostin AG 879, DMPQ, JNJ-10198409, PLX647, Trapidil,Tyrphostin A9, Tyrphostin AG 370, Lestaurtinib, DMH4, Geldanamycin,Genistein, GW2580, Herbimycin A, Lavendustin C, Midostaurin, NVP-BHG712,PD158780, PD-166866, PF-06273340, PP2, RPI, SU 11274, SU5614, Symadex,Tyrphostin AG 34, Tyrphostin AG 974, Tyrphostin AG 1007, UNC2881,Honokiol, SU1498, SKLB1002, CP-547632, JK-P3, KRN633, SC-1, ST638, SU5416, Sulochrin, Tyrphostin SU 1498, S8567, rociletinib, Dacomitinib,Tivantinib, Neratinib, Masitinib, Vatalanib, Icotinib, XL-184, OSI-930,AB1010, Quizartinib, AZD9291, Tandutinib, HM61713, Brigantinib,Vemurafenib (PLX-4032), Semaxanib, AZD2171, Crenolanib, Damnacanthal,Fostamatinib, Motesanib, Radotinib, OSI-027, Linsitinib, BIX02189,PF-431396, PND-1186, PF-03814735, PF-431396, sirolimus, temsirolimus,everolimus, deforolimus, zotarolimus, BEZ235, INK128, Omipalisib,AZD8055, MHY1485, PI-103, KU-0063794, ETP-46464, GDC-0349, XL388,WYE-354, WYE-132, GSK1059615, WAY-600, PF-04691502, WYE-687, PP121,BGT226, AZD2014, PP242, CH5132799, P529, GDC-0980, GDC-0994, XMD8-92,Ulixertinib, FR180204, SCH772984, Trametinib, PD184352, PD98059,Selumetinib, PD325901, U0126, Pimasertinib, TAK-733, AZD8330,Binimetinib, PD318088, SL-327, Refametinib, GDC-0623, Cobimetinib,BI-847325, Adaphostin, GNF 2, PPY A, AIM-100, ASP 3026, LFM A13, PF06465469, (−)-Terreic acid, AG-490, BIBU 1361, BIBX 1382, BMS 599626,CGP 52411, GW 583340, HDS 029, HKI 357, JNJ 28871063, WHI-P 154, PF431396, PF 573228, FUN 1, PD 166285, SUN 11602, SR 140333, TCS 359, BMS536924, NVP ADW 742, PQ 401, BMS 509744, CP 690550, NSC 33994, WHI-P154, KB SRC 4, DDR1-IN-1, PF 04217903, PHA 665752, SU 16f, A 419259, AZM475271, PP 1, PP 2, 1-Naphthyl PP1, Src I1, ANA 12, PD 90780, Ki 8751,Ki 20227, ZM 306416, ZM 323881, AEE 788, GTP 14564, PD 180970, R 1530,SU 6668, and Toceranib.

In certain embodiments, the TKI of the invention is selected from thegroup consisting of CEP-32496(1-(3-((6,7-dimethoxyquinazolin-4-yl)oxy)phenyl)-3-(5-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazol-3-yl)urea),AZ628(4-(2-cyanopropan-2-yl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)benzamide),Vemurafenib (PLX-4032), PLX-4720(N-(3-(5-chloro-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide),SB 590885((E)-5-(2-(4-(2-(dimethylamino)ethoxy)phenyl)-4-(pyridin-4-yl)-1H-imidazol-5-yl)-2,3-dihydro-1H-inden-1-oneoxime), andGDC-0879((E)-5-(2-(2-hydroxyethyl)-4-(pyridin-4-yl)-1H-imidazol-5-yl)-2,3-dihydro-1H-inden-1-oneoxime).

In certain embodiments, the TKI of the invention is

wherein R¹ is H or CH₃, and R² is

In certain embodiments, the TKI of the invention is

wherein R is

In certain embodiments, the TKI of the invention is

wherein the broken lines correspond to the divalent group

Bosutinib is also known as4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-methyl group is replaced with L.Exemplary positions wherein L may be attached are illustrated below:

Dasatinib is also known asN-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-(2-hydroxyethyl) group is derivatizedwith and/or replaced with L. Exemplary positions wherein L may beattached are illustrated below:

Imatinib is also known asN-(4-methyl-3-(4-(pyridin-3-yl)pyrimidin-2-ylamino)phenyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-methyl group is replaced with L.Exemplary positions wherein L may be attached are illustrated below:

Saracatinib is also known asN-(5-chlorobenzo[d][1,3]dioxol-4-yl)-7-(2-(4-methylpiperazin-1-yl)ethoxy)-5-(tetrahydro-2H-pyran-4-yloxy)quinazolin-4-amine,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-methyl group is replaced with L.Exemplary positions wherein L may be attached are illustrated below:

Ponatinib is also known as3-(2-(imidazo[1,2-b]pyridazin-3-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)benzamide,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-methyl group is replaced with L.Exemplary positions wherein L may be attached are illustrated below:

Nilotinib is also known as4-methyl-N-(3-(4-methyl-1H-imidazol-1-yl)-5-(trifluoromethyl)phenyl)-3-(4-(pyridin-3-yl)pyrimidin-2-ylamino)benzamide,and has a formula of:

Exemplary positions wherein L may be attached are illustrated below:

Danusertib is also known as(R)—N-(5-(2-methoxy-2-phenylacetyl)-1,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-methyl group is replaced with L.Exemplary positions wherein L may be attached are illustrated below:

AT9283 is also known as1-cyclopropyl-3-(3-(5-(morpholinomethyl)-1H-benzo[d]imidazol-2-yl)-1H-pyrazol-4-yl)urea,and has a formula of:

Linker L can be connected, for example, to the morpholino ring of thisTKI. Exemplary positions wherein L may be attached are illustratedbelow:

Degrasyn is also known as(S,E)-3-(6-bromopyridin-2-yl)-2-cyano-N-(1-phenyl butyl)acrylamide, andhas a formula of:

Exemplary positions wherein L may be attached are illustrated below:

Bafetinib is also known as(S)-4-((3-(dimethylamino)pyrrolidin-1-yl)methyl)-N-(4-methyl-3-(4-(pyrimidin-5-yl)pyrimidin-2-ylamino)phenyl)-3-(trifluoromethyl)benzamide,and has a formula of:

L can be connected, for example, to the pyrrolidine ring of this TKI. Incertain embodiments, at least one of the N-methyl groups is replacedwith L. Exemplary positions wherein L may be attached are illustratedbelow:

KW-2449 is also known as(E)-(4-(2-(1H-indazol-3-yl)vinyl)phenyl)(piperazin-1-yl)methanone, andhas a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the NH— group of the piperidine group islinked to L. Exemplary positions wherein L may be attached areillustrated below:

NVP-BHG712 is also known as4-methyl-3-(1-methyl-6-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino)-N-(3-(trifluoromethyl)phenyl)benzamide,and has a formula of:

Exemplary positions wherein L may be attached are illustrated below:

DCC-2036 is also known as1-(3-tert-butyl-1-(quinolin-6-yl)-1H-pyrazol-5-yl)-3-(2-fluoro-4-(2-(methylcarbamoyl)pyridin-4-yloxy)phenyl)urea,and has a formula of:

Exemplary positions wherein L may be attached are illustrated below:

GZD824 is also known as4-methyl-N-[4-[(4-methyl-1-piperazinyl)methyl]-3-(trifluoromethyl)phenyl]-3-[2-(1H-pyrazolo[3,4-b]pyridin-5-yl)ethynyl]-benzamide,and has a formula of:

Linker L can be connected, for example, to the piperidine ring of thisTKI. In certain embodiments, the N-methyl group is replaced with L.Exemplary positions wherein L may be attached are illustrated below:

GNF-2 is also known as3-[6-[[4-(trifluoromethoxy)phenyl]amino]-4-pyrimidinyl]-benzamide, andhas a formula of:

Exemplary positions wherein L may be attached are illustrated below:

PD173955 is also known as6-(2,6-dichlorophenyl)-8-methyl-2-((3-(methylthio)phenyl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one,and has a formula of:

Exemplary positions wherein L may be attached are illustrated below:

GNF-5 is also known as N-(2-Hydroxyethyl)-3-[6-[[4-(trifluoromethoxy)phenyl]amino]-4-pyrimidinyl]benzamide, and has a formula of:

Linker L can be connected, for example, to the terminal amide of thisTKI In certain embodiments, the N-2-hydroxyethyl group is derivatizedwith and/or replaced with L. Exemplary positions wherein L may beattached are illustrated below:

Gefitinib is also known asN-(3-chloro-4-fluoro-phenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine,and has a formula of:

Linker L can be connected, for example, to the morpholino ring of thisTKI. Exemplary positions wherein L may be attached are illustratedbelow:

Erlotinib is also known as N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine, and has a formula of:

Linker L can be connected, for example, to the ether chains of this TKI.In certain embodiments, at least one of the O-methyl groups is replacedwith L. Exemplary positions wherein L may be attached are illustratedbelow:

Sunitinib is also known asN-(2-diethylaminoethyl)-5-[(Z)-(5-fluoro-2-oxo-1H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide,and has a formula of:

Linker L can be connected, for example, to the amine group of this TKI.In certain embodiments, at least one of the n-ethyl groups is replacedwith L. Exemplary positions wherein L may be attached are illustratedbelow:

Vemurafenib is also known asN-(3-(5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-carbonyl)-2,4-difluorophenyl)propane-1-sulfonamide,and has a formula of

Linker can be connected, for example, to the compound as illustratedherein:

Linker (L)

A suitable linker of the present invention is covalently bonded to theTKI, and is further covalently bonded to at least one ubiquitin ligasebinding. In certain embodiment, the ubiquitin ligase is an E3 ubiquitinligase. In other embodiments, the ubiquitin ligase is Von Hippel Lindau(VHL) E3 ubiquitin ligase and/or Cereblon (CRBN) E3 ligase.

In certain embodiments, the linker of the present invention is a bond.

In certain embodiments, the linker of the present invention correspondsto formula —(CH₂)_(m1)—X₄—(CH₂—CH₂—X₅)_(m2)—(CH₂)_(m3)—C(X₆)—, whereinthe TKI is covalently bonded to —(CH₂)_(m1), and the ULM is covalentlybonded to C(X₆)—. Alternatively, —(CH₂)_(m1) is covalently bonded to theULM, and C(X₆)— is covalently bonded to the TKI. Each m1, m2, and m3 isindependently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each X₄, X₅, and X₆is independently absent (a bond), O, S, or N—R²⁰, wherein each R²⁰ isindependently selected from the group consisting of hydrogen, optionallysubstituted C₁-C₆ alkyl, optionally substituted aryl, optionallysubstituted heteroaryl, optionally substituted C₃-C₈ cycloalkyl, andoptionally substituted C₃-C₈ cycloheteroalkyl.

In other embodiments, the linker of the present invention corresponds toformula —(CH₂)_(m1)—O—(CH₂—CH₂—O)_(m2)—(CH₂)_(m3)—C(O)—, wherein the TKIis covalently bonded to —(CH₂)_(m1), and the ULM is covalently bonded toC(O)—. Alternatively, —(CH₂)_(m1) is covalently bonded to the ULM, andC(O)— is covalently bonded to the TKI. Each m1, m2, and m3 is definedelsewhere herein.

In yet other embodiments, the linker of the present inventioncorresponds to formula—(CHR₂₁)_(m1)—O—(CHR₂₂—CHR₂₃—O)_(m2)—(CHR₂₄)_(m3)—C(O)—, wherein the TKIis covalently bonded to —(CH₂)_(m1), and the ULM is covalently bonded toC(O)—. Alternatively, —(CH₂)_(m1) is covalently bonded to the ULM, andC(O)— is covalently bonded to the TKI. Each m1, m2, and m3 is definedelsewhere herein; each R₂₁, R₂₂, R₂₃, and R₂₄ is independently selectedfrom the group consisting of hydrogen, optionally substituted C₁-C₆alkyl, optionally substituted aryl, optionally substituted heteroaryl,optionally substituted C₃-C₈ cycloalkyl, and optionally substitutedC₃-C₈ cycloheteroalkyl.

In yet other embodiments, L is a polyethylene glycol chain ranging insize from about 1 to about 12 ethylene glycol units, from about 1 toabout 10 ethylene glycol units, from about 2 to about 6 ethylene glycolunits, from about 2 to about 5 ethylene glycol units, or from about 2 toabout 4 ethylene glycol units.

In yet other embodiments, the linker L corresponds to -(D-CON-D)_(m1)-(II), wherein each D is independently a bond (absent), or—(CH₂)_(m1)—Y—C(O)—Y—(CH₂)_(m1)—; wherein m1 is defined elsewhereherein; Y is O, S or N—R⁴; CON is a bond (absent), an optionallysubstituted C₃—C cycloheteroalkyl, piperazinyl or a group selected fromthe group consisting of the following chemical structures:

wherein X² is O, S, NR⁴, S(O), S(O)₂, —S(O)₂O, —OS(O)₂, or OS(O)₂O; X³is O, S, CHR⁴, NR⁴; and R⁴ is H or a C₁-C₃ alkyl group optionallysubstituted with one or two hydroxyl groups.

The linker L of the present invention is covalently bonded to the TKIand ULM, suitable through an amide, ester, thioester, keto group,carbamate (urethane) or ether. The linking position can be anywhere inthe TKI and ULM. One of ordinary skill in the art would recognize thesuitable linking positions to maximum the binding affinity between theTKI and tyrosine kinase, and between the ULM and the ubiquitin ligase.

Ubiquitin Ligase Moiety (ULM)

A ubiquitin ligase binder (ULM) of a compound of the present inventionbinds to a ubiquitin ligase. In certain embodiments, the ubiquitinligase is an E3 ubiquitin ligase.

In certain embodiments, ULM corresponds to formula (III):

wherein

R^(1′) is a group selected from the group consisting of an optionallysubstituted C₁-C₆ alkyl, an optionally substituted —(CH₂)_(n)OH, anoptionally substituted —(CH₂)_(n)SH, an optionally substituted(CH₂)_(n)—O—(C₁-C₆)alkyl, an optionally substituted(CH₂)_(n)—X₇—(C₁-C₆)alkyl, an optionally substituted —(CH₂)_(n)COOH, anoptionally substituted —(CH₂)_(n)C(O)—(C₁-C₆ alkyl), an optionallysubstituted —(CH₂)_(n)NHC(O)—R₁, an optionally substituted—(CH₂)_(n)C(O)—NR₁R₂, an optionally substituted —(CH₂)_(n)OC(O)—NR₁R₂,—(CH₂O)_(n)H, an optionally substituted —(CH₂)_(n)OC(O)—(C₁-C₆ alkyl),an optionally substituted —(CH₂)_(n)C(O)—O—(C₁-C₆ alkyl), an optionallysubstituted —(CH₂O)_(n)COOH, an optionally substituted—(OCH₂)_(n)O—(C₁-C₆ alkyl), an optionally substituted—(CH₂O)_(n)C(O)—(C₁-C₆ alkyl), an optionally substituted—(OCH₂)_(n)NHC(O)—R₁, an optionally substituted —(CH₂O)_(n)C(O)—NR₁R₂,—(CH₂CH₂O)_(n)H, an optionally substituted —(CH₂CH₂O)_(n)COOH, anoptionally substituted —(OCH₂CH₂)_(n)O—(C₁-C₆ alkyl), an optionallysubstituted —(CH₂CH₂O)_(n)C(O)—(C₁-C₆ alkyl), an optionally substituted—(OCH₂CH₂)_(n)NHC(O)—R₁, an optionally substituted—(CH₂CH₂O)_(n)C(O)—NR₁R₂, an optionally substituted —SO₂R_(S),S(O)R_(S), NO₂, CN, and halogen;

R₁ and R₂ are each independently H or C₁-C₆ alkyl which may beoptionally substituted with one or two hydroxyl groups or up to threehalogen groups;

R_(S) is C₁-C₆ alkyl, optionally substituted aryl, optionallysubstituted heteroaryl or optionally substituted heterocycle or—(CH₂)_(m)NR₁R₂;

X and X′ are each independently C═O, C═S, —S(O), S(O)₂;

X₇ is an optionally substituted epoxide moiety;

R² is a group selected from the group consisting of optionallysubstituted —(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—C₁-C₆ alkyl, anoptionally substituted—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl, an optionallysubstituted —(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl, anoptionally substituted —(CH₂)_(n)—(C═O)_(v)NR(SO₂)_(w)-Heterocycle, anoptionally substituted —NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—C₁-C₆alkyl, an optionally substituted—NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), anoptionally substituted —NR₂₅—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl,an optionally substituted—NR₂₅—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl or an optionallysubstituted —NR₂₅—(CH₂)_(n)—(C═O)_(v)NR(SO₂)_(w)-Heterocycle, anoptionally substituted —X^(R2′)—C₁-C₆ alkyl; an optionally substituted—X^(R2′)-Aryl, an optionally substituted —X^(R2′)-Heteroaryl, and anoptionally substituted —X^(R2′)-Heterocycle;

R³ is a group selected from the group consisting of an optionallysubstituted C₁-C₆ alkyl, an optionally substituted—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—C₁-C₆ alkyl, an optionallysubstituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), anoptionally substituted—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), an optionallysubstituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—C(O)NR₁R₂, anoptionally substituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl, anoptionally substituted —(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl,an optionally substituted—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Heterocycle, an optionallysubstituted —NR₂₅—(CH₂)_(n)—C(O)₁₁(NR₁)_(v)(SO₂)_(w)—C₁-C₆ alkyl, anoptionally substituted—NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), anoptionally substituted —NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl,an optionally substituted—NR₂₅—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl, an optionallysubstituted —NR_(1′)—(CH₂)_(n)—C(O)_(u)(NR₁)_(v)(SO₂)_(w)-Heterocycle,an optionally substituted —O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—C₁-C₆alkyl, an optionally substituted—O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—NR_(1N)R_(2N), an optionallysubstituted —O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)—NR₁C(O)R_(1N), anoptionally substituted —O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Aryl, anoptionally substituted—O—(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Heteroaryl, an optionallysubstituted —(CH₂)_(n)—(C═O)_(u)(NR₁)_(v)(SO₂)_(w)-Heterocycle,—(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)—C₁-C₆ alkyl, an optionallysubstituted —(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)-Aryl, an optionallysubstituted —(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)-Heteroaryl, anoptionally substituted—(CH₂)_(n)—(V)_(n′)—(CH₂)_(n)—(V)_(n′)-Heterocycle, an optionallysubstituted —(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)—C₁-C₆ alkyl, anoptionally substituted —(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-Aryl, anoptionally substituted —(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-Heteroaryl,an optionally substituted—(CH₂)_(n)—N(R₁)(C═O)_(m′)—(V)_(n′)-Heterocycle, an optionallysubstituted —X^(R3′)—C₁-C₆ alkyl group; an optionally substituted—X^(R3′)-Aryl group; an optionally substituted —X^(R3′)-Heteroarylgroup; and an optionally substituted —X^(R3′)-Heterocycle group;

where R_(1N) and R_(2N) are each independently H, C₁-C₆ alkyl which isoptionally substituted with one or two hydroxyl groups and up to threehalogen groups or an optionally substituted —(CH₂)_(n)-Aryl,—(CH₂)_(n)-Heteroaryl or —(CH₂)_(n)-Heterocycle group;

V is O, S or NR₁;

each R₂₅ is independently H or C₁-C₃ alkyl;

X^(R2′) and X^(R3′) are each independently an optionally substituted—CH₂)_(n)—, —CH₂)_(n)—CH(X_(v))═CH(X_(v))— (cis or trans),—CH₂)_(n)—CH—CH—, —(CH₂CH₂O)_(n)— or a C₃-C₆ cycloalkyl, where X_(v) isH, a halo or optionally substituted C₁-C₃ alkyl;

Each m is independently 0, 1, 2, 3, 4, 5, 6;

Each m′ is independently 0 or 1;

Each n is independently 0, 1, 2, 3, 4, 5, 6;

Each n′ is independently 0 or 1;

Each u is independently 0 or 1;

Each v is independently 0 or 1;

Each w is independently 0 or 1; and

wherein any one or more of R^(1′), R^(2′), R^(3′), X and X′ of ULM groupis modified to be covalently bonded to the TKI group through a linker L.

In other embodiments, the ULM corresponds to formula (IV) or (V):

wherein each of X, R^(1′), R^(2′) and R^(3′) are defined elsewhereherein.

In certain embodiments of the formulas (III), (IV), and (V), R^(1′) is ahydroxyl group or a group which may be metabolized to a hydroxyl orcarboxylic group. Exemplary R^(1′) groups include —(CH₂)_(n)OH,—(CH₂)_(n)—O—(C₁-C₆)alkyl, —(CH₂)_(n)COOH, —(CH₂O)_(n)H, an optionallysubstituted —(CH₂)_(n)OC(O)—(C₁-C₆ alkyl), or an optionally substituted—(CH₂)_(n)C(O)—O—(C₁-C₆ alkyl), wherein n is defined above.

In yet other embodiments of the formulas (III), (IV), and (V), R² andR^(3′) are each independently selected from the group consisting of anoptionally substituted —NR₂₆-T-Aryl, an optionally substituted—NR₂₆-T-Heteroaryl or an optionally substituted —NR₂₆-T-Heterocycle,wherein R₂₆ is H or CH₃, and T is a group selected from the groupconsisting of —(CH₂)_(n)—, —(CH₂O)_(n)—, —(OCH₂)_(n)—, —(CH₂CH₂O)—, and—(OCH₂CH₂)—, wherein each one of the methylene groups may be optionallysubstituted with one or two substituents, selected from the groupconsisting of halogen, an amino acid, and C₁-C₃ alkyl; wherein n isdefined above.

In yet other embodiments of the formulas (III), (IV), and (V), R^(2′) orR³ is —NR₂₆-T-Ar¹, wherein the Ar¹ is phenyl or naphthyl optionallysubstituted with a group selected from the group consisting of a linkergroup L to which is attached a TKI moiety, a halogen, an amine,monoalkyl- or dialkyl amine (preferably, dimethylamine), OH, COOH, C₁-C₆alkyl, CF₃, OMe, OCF₃, NO₂, CN, an optionally substituted phenyl, anoptionally substituted naphthyl, and an optionally substitutedheteroaryl. Suitable heteroaryl includes an optionally substitutedisoxazole, an optionally substituted oxazole, an optionally substitutedthiazole, an optionally substituted isothiazole, an optionallysubstituted pyrrole, an optionally substituted imidazole, an optionallysubstituted benzimidazole, an optionally substituted oximidazole, anoptionally substituted diazole, an optionally substituted triazole, anoptionally substituted pyridine or an oxapyridine, an optionallysubstituted furan, an optionally substituted benzofuran, an optionallysubstituted dihydrobenzofuran, an optionally substituted indole,indolizine, azaindolizine, an optionally substituted quinoline, and anoptionally substituted group selected from the group consisting of thechemical structures:

wherein S^(c) is CHR^(SS), NR^(URE), or O; R^(HET) is H, CN, NO₂, halo,optionally substituted C₁-C₆ alkyl, optionally substituted O(C₁-C₆alkyl) or an optionally substituted acetylenic group —C≡C—R_(a), whereinR_(a) is H or C₁-C₆ alkyl; R^(SS) is H, CN, NO₂, halo, optionallysubstituted C₁-C₆ alkyl, optionally substituted O—(C₁-C₆ alkyl oroptionally substituted —C(O)(C₁-C₆ alkyl); R^(URE) is H, C₁-C₆ alkyl or—C(O)(C₁-C₆ alkyl), wherein the alky group is optionally substitutedwith one or two hydroxyl groups, up to three halogens, an optionallysubstituted phenyl group, an optionally substituted heteroaryl, or anoptionally substituted heterocycle, preferably for example piperidine,morpholine, pyrrolidine, tetrahydrofuran; R^(PRO) is H, optionallysubstituted C₁-C₆ alkyl, an optionally substituted aryl, an optionallysubstituted heteroaryl or an optionally substituted heterocyclic groupselected from the group consisting of oxazole, isoxazole, thiazole,isothiazole, imidazole, diazole, oximidazole, pyrrole, pyrollidine,furan, dihydrofuran, tetrahydrofuran, thiene, dihydrothiene,tetrahydrothiene, pyridine, piperidine, piperazine, morpholine,quinoline, benzofuran, indole, indolizine, and azaindolizine; R^(PRO1)and R^(PRO2) are each independently H, optionally substituted C₁-C₃alkyl or together form a keto group; n is defined above.

In yet other embodiments of the formulas (III), (IV), and (V), R^(2′) orR^(3′) is an optionally substituted —NR₂₆-T-Ar² group, wherein the Ar²group is selected from the group consisting of quinoline, indole,indolizine, azaindolizine, benzofuran, isoxazole, thiazole, isothiazole,thiophene, pyridine, imidazole, pyrrole, diazole, triazole, tetrazole,oximidazole, and a group selected from the group consisting of thefollowing chemical structures:

wherein S^(c), R^(HET), and R^(URE) are defined elsewhere herein; Y^(C)is N or C—R^(YC); R^(YC) is H, OH, CN, NO₂, halo, optionally substitutedC₁-C₆ alkyl, optionally substituted O(C₁-C₆ alkyl), or an optionallysubstituted acetylenic group —C≡C—R_(a); R_(a) is H or C₁-C₆ alkyl.

In yet other embodiments of the formulas (III), (IV), and (V), R^(2′) orR^(3′) is an optionally substituted —NR₂₆-T-HET¹, wherein the HET¹ isselected from the group consisting of tetrahydrofuran, tetrahydrothiene,tetrahydroquinoline, piperidine, piperazine, pyrrollidine, morpholine,oxane and thiane. The HET¹ is optionally substituted by a group selectedfrom the group consisting of the following chemical structures:

wherein n, R^(PRO), R^(PRO1), R^(HET) and R^(PRO2) are defined elsewhereherein.

In other embodiments, R^(2′) or R^(3′) is optionally substituted—(CH₂)_(n)—(V)_(n)—(CH₂)_(n)—(V)_(n′)—R^(S3′), optionally substituted—(CH₂)_(n)—N(R₂₆)(C═O)_(m′)—(V)_(n′)—R^(S3′), optionally substituted—X^(R3′)—C₁-C₁₀ alkyl, optionally substituted —X^(R3′)—Ar³, optionallysubstituted —X^(R3′)-HET, optionally substituted —X^(R3′)—Ar³-HET oroptionally substituted —X^(R3′)-HET-Ar³, wherein R^(S3′) is optionallysubstituted C₁-C₁₀ alkyl, optionally substituted Ar³ or HET; R₂₆ isdefined elsewhere herein; V is O, S or NR_(1′); X^(R3′) is —(CH₂)_(n)—,—(CH₂CH₂O)_(n)—, —CH₂)_(n)—CH(X_(v))═CH(X_(v))— (cis or trans),—CH₂)_(n)-CH—CH—, or a C₃-C₆ cycloalkyl group, all optionallysubstituted; wherein X, is H, a halo or a C₁-C₃ alkyl group which isoptionally substituted with one or two hydroxyl groups or up to threehalogen groups; Ar³ is an optionally substituted phenyl or napthylgroup; and HET is an optionally substituted oxazole, isoxazole,thiazole, isothiazole, imidazole, diazole, oximidazole, pyrrole,pyrollidine, furan, dihydrofuran, tetrahydrofuran, thiene,dihydrothiene, tetrahydrothiene, pyridine, piperidine, piperazine,morpholine, benzofuran, indole, indolizine, azaindolizine, quinoline, ora group selected from the group consisting of the following chemicalstructures:

wherein n, v, n′, m′, S^(c), R^(HET), R^(URE), Y^(C), R^(PRO1) andR^(PRO2) are defined elsewhere herein.

In yet other embodiments of the formulas (III), (IV), and (V), R^(2′) orR^(3′) is an optionally substituted —NR₂₆—X^(R2′)—C₁-C₁₀ alkyl,—NR₂₆—X^(R2′)—Ar³, an optionally substituted —NR₂₆—X^(R2′)-HET, anoptionally substituted —NR₂₆—X^(R2′)—Ar³-HET, or an optionallysubstituted —NR₂₆—X^(R2′)-HET-Ar³, X^(R2′) is an optionally substituted—CH₂)_(n)—, —CH₂)_(n)—CH(X_(v))═CH(X_(v))— (cis or trans),—CH₂)_(n)—CH≡CH—, —(CH₂CH₂O)_(n)— or C₃-C₆ cycloalkyl; wherein X, is H,a halo or a C₁-C₃ alkyl group which is optionally substituted with oneor two hydroxyl groups or up to three halogen groups; wherein HET, Ar³,and R₂₆ are defined elsewhere herein.

In yet other embodiments, R^(2′) or R^(3′) is —(CH₂)_(n)—Ar¹,—(CH₂CH₂O)—Ar¹, —(CH₂)_(n)-HET or —(CH₂CH₂O)_(n)-HET; wherein n, Ar¹,and HET are defined elsewhere herein.

In yet other embodiments, ULM corresponds to formula (VI):

wherein R^(1′) is OH or a group which is metabolized in a patient orsubject to OH; R^(2′) is —NH—CH₂—Ar⁴-HET¹; R^(3′) is—CHR^(CR3′)—NH—C(O)—R^(3P1) or —CHR^(CR3′)R^(3P2); wherein R^(CR3′) icC₁-C₄ alkyl, preferably methyl, isopropyl or tert-butyl; R^(3P1) isC₁-C₃ alkyl, optionally substituted oxetane, —CH₂OCH₃, —CH₂CH₂OCH₃,morpholino, or

R^(3P2) is a

group, wherein Ar⁴ is phenyl; HET¹ is an optionally substituted thiazoleor isothiazole; and R^(HET) is H or halo.

In yet other embodiments, ULM corresponds to formula (VII) or (VIII):

wherein X₅ is Cl, F, C₁-C₃ alkyl or heterocycle; R₂₇ and R₂₈ are eachindependently H, C₁-C₃ alkyl.

In yet other embodiments, ULM is a cereblon ligand of formula (IX) or aVHL ligand of formula (X):

In certain embodiments, the compound of the present invention is acompound of formula (XI):

wherein n¹ is 0 or 1; X₅ is H, F, Cl, C₁-C₃ alkyl or heterocycle.

In yet other embodiments, the compound of the present invention is acompound of formula (XII):

wherein n and X₅ are defined elsewhere herein; R₂₉ is C₁-C₃ alkyl or—C(O)NR₃₀R₃₁ where R₃₀ and R₃₁ are each independently H, C₁-C₃ alkyl,phenyl or heterocycle.

In yet other embodiments, the compound of the present invention is acompound of formula (XIII):

wherein n¹ and R₂₉ are defined elsewhere herein.

In yet other embodiments, the compound of the present invention is acompound of formula (XIV):

wherein n¹ is defined elsewhere herein; each X₆ is independently is H,F, Cl, C₁-C₃ alkyl, heterocycle, —O—C(O)NR₃₂R₃₃ or —C(O)NR₃₂R₃₃, whereineach of R₃₂ and R₃₃ is independently H, C₁-C₃ alkyl, or phenyl.

In yet other embodiments, the compound of the present invention is acompound of formula (XV):

wherein n, R₂₉, and X₅ are defined elsewhere herein.

In yet other embodiments, the compound of the present invention is acompound of formula (XVI):

wherein n, R₂₉, and X₅ are defined elsewhere herein.

In yet other embodiments, the compound of the present invention is acompound of formula (XVII):

wherein either of R_(7PC) or R_(10PC) is a -L-TKI group and the otherR_(7PC) or R_(10PC) is H.

In yet other embodiments, the compound of the present invention is acompound of formula (XVIII):

wherein R_(7PC) is a -L-TKI group.

In yet other embodiments, the compound of the present invention is acompound of formula (XIX):

wherein R_(7PC), R_(11PC), R_(12PC), R_(13PC) and R_(14PC) are eachindependently a -L-TKI group or H; one of R_(7PC), R_(11PC), R_(12PC),R_(13PC) and R_(14PC) is a -L-TKI group and the other groups are H.

In yet other embodiments, the compound of the present invention is acompound of formula (XX):

wherein R_(4PC), R_(7PC), R_(11PC), R_(12PC), R_(13PC), and R_(14PC) areeach independently a -L-TKI group or H; either of R_(4PC), R_(7PC) orone of R_(11PC), R_(12PC), R_(13PC) and R_(14PC) is a -L-TKI group andthe other groups are H.

In yet other embodiments, the compound of the present invention is acompound of formula (XXI):

wherein R_(3PC), R_(7PC), R_(11PC), R_(12PC), R_(13PC) and R_(14PC) areeach independently a -L-TKI group or H; one of R_(3PC), R_(7PC),R_(11PC), R_(12PC), R_(13PC) and R_(14PC) is a -L-TKI group and theother groups are H.

In yet other embodiments, the compound of the present invention is acompound of formula (XXII):

wherein R_(7PC) and R_(10PC) are each independently a -L-TKI group or H;one of R_(7PC) and R_(10PC) is a -L-TKI group and the other group is H.

In yet other embodiments, the compound of the present invention is acompound of formula (XXIII), (XXIV), (XXV), or (XXVI):

wherein R_(7PC) and R_(10RC) are each independently a -L-TKI group or Hand R_(8PC) is H or CH₃. In one embodiment, one of R_(7PC) and R_(10PC)is a -L-TKI group and the other group is H and R_(8PC) is H.

In yet other embodiments, the compound of the present invention is acompound of formula of (XXVI):

wherein R_(7PC) and R_(9PC) are each independently a -L-TKI group or H.In one embodiment, one of R_(7PC) and R_(9PC) is a -L-TKI group and theother group is H.

In yet other embodiments, the compound of the present invention is acompound of formula (XXVIII):

wherein R_(7PC) and R_(14PC) are each independently a -L-TKI group or Hand each of R_(12PC) and R_(13PC) is H or CH₃. In one embodiment, one ofR_(7PC) and R_(14PC) is a -L-TKI group and the other of R_(7PC) andR_(14PC) group is H and each of R_(12PC) and R_(13PC) is H.

In yet other embodiments, the compound of the present invention is acompound of formula (XXIX) or (XXX):

wherein R_(7PC) and R_(9PC) are each independently a -L-TKI group or H.In one embodiment, one of R_(7PC) and R_(9PC) is a -L-TKI group and theother group is H.

In yet other embodiments, the compound of the present invention is acompound of formula (XXXI):

wherein R_(7PC) and R_(10PC) are each independently a -L-TKI group or Hand R_(9PC) is H or CH₃. In one embodiment, one of R_(7PC) and R_(10PC)is a -L-TKI group and the other group is H and R_(9PC) is H.

Preparation of Compounds of the Invention

Compounds of formulas (I)-(XXXI) may be prepared by the general schemesdescribed herein, using the synthetic method known by those skilled inthe art. The following examples illustrate non-limiting embodiments ofthe invention.

The compounds of the invention may possess one or more stereocenters,and each stereocenter may exist independently in either the (R) or (S)configuration. In certain embodiments, compounds described herein arepresent in optically active or racemic forms. It is to be understoodthat the compounds described herein encompass racemic, optically-active,regioisomeric and stereoisomeric forms, or combinations thereof thatpossess the therapeutically useful properties described herein.Preparation of optically active forms is achieved in any suitablemanner, including by way of non-limiting example, by resolution of theracemic form with recrystallization techniques, synthesis fromoptically-active starting materials, chiral synthesis, orchromatographic separation using a chiral stationary phase. In certainembodiments, a mixture of one or more isomer is utilized as thetherapeutic compound described herein. In other embodiments, compoundsdescribed herein contain one or more chiral centers. These compounds areprepared by any means, including stereoselective synthesis,enantioselective synthesis and/or separation of a mixture of enantiomersand/or diastereomers. Resolution of compounds and isomers thereof isachieved by any means including, by way of non-limiting example,chemical processes, enzymatic processes, fractional crystallization,distillation, and chromatography.

The methods and formulations described herein include the use ofN-oxides (if appropriate), crystalline forms (also known as polymorphs),solvates, amorphous phases, and/or pharmaceutically acceptable salts ofcompounds having the structure of any compound of the invention, as wellas metabolites and active metabolites of these compounds having the sametype of activity. Solvates include water, ether (e.g., tetrahydrofuran,methyl tert-butyl ether) or alcohol (e.g., ethanol) solvates, acetatesand the like. In certain embodiments, the compounds described hereinexist in solvated forms with pharmaceutically acceptable solvents suchas water, and ethanol. In other embodiments, the compounds describedherein exist in unsolvated form.

In certain embodiments, the compounds of the invention may exist astautomers. All tautomers are included within the scope of the compoundspresented herein.

In certain embodiments, compounds described herein are prepared asprodrugs. A “prodrug” refers to an agent that is converted into theparent drug in vivo. In certain embodiments, upon in vivoadministration, a prodrug is chemically converted to the biologically,pharmaceutically or therapeutically active form of the compound. Inother embodiments, a prodrug is enzymatically metabolized by one or moresteps or processes to the biologically, pharmaceutically ortherapeutically active form of the compound.

In certain embodiments, sites on, for example, the aromatic ring portionof compounds of the invention are susceptible to various metabolicreactions. Incorporation of appropriate substituents on the aromaticring structures may reduce, minimize or eliminate this metabolicpathway. In certain embodiments, the appropriate substituent to decreaseor eliminate the susceptibility of the aromatic ring to metabolicreactions is, by way of example only, a deuterium, a halogen, or analkyl group.

Compounds described herein also include isotopically-labeled compoundswherein one or more atoms is replaced by an atom having the same atomicnumber, but an atomic mass or mass number different from the atomic massor mass number usually found in nature. Examples of isotopes suitablefor inclusion in the compounds described herein include and are notlimited to ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ³⁶Cl, ¹⁸F, ¹²³I, ¹²⁵I, ¹³N, ¹⁵N, ¹⁵O,¹⁷O, ¹⁸O, ³²P, and ³⁵S. In certain embodiments, isotopically-labeledcompounds are useful in drug and/or substrate tissue distributionstudies. In other embodiments, substitution with heavier isotopes suchas deuterium affords greater metabolic stability (for example, increasedin vivo half-life or reduced dosage requirements). In yet otherembodiments, substitution with positron emitting isotopes, such as ¹¹C,¹⁸F, ¹⁵O and ¹³N, is useful in Positron Emission Topography (PET)studies for examining substrate receptor occupancy. Isotopically-labeledcompounds are prepared by any suitable method or by processes using anappropriate isotopically-labeled reagent in place of the non-labeledreagent otherwise employed.

In certain embodiments, the compounds described herein are labeled byother means, including, but not limited to, the use of chromophores orfluorescent moieties, bioluminescent labels, or chemiluminescent labels.

The compounds described herein, and other related compounds havingdifferent substituents are synthesized using techniques and materialsdescribed herein and as described, for example, in Fieser & Fieser'sReagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons,1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 andSupplementals (Elsevier Science Publishers, 1989); Organic Reactions,Volumes 1-40 (John Wiley and Sons, 1991), Larock's Comprehensive OrganicTransformations (VCH Publishers Inc., 1989), March, Advanced OrganicChemistry 4^(th) Ed., (Wiley 1992); Carey & Sundberg, Advanced OrganicChemistry 4th Ed., Vols. A and B (Plenum 2000,2001), and Green & Wuts,Protective Groups in Organic Synthesis 3rd Ed., (Wiley 1999) (all ofwhich are incorporated by reference for such disclosure). Generalmethods for the preparation of compound as described herein are modifiedby the use of appropriate reagents and conditions, for the introductionof the various moieties found in the formula as provided herein.

Compounds described herein are synthesized using any suitable proceduresstarting from compounds that are available from commercial sources, orare prepared using procedures described herein.

In certain embodiments, reactive functional groups, such as hydroxyl,amino, imino, thio or carboxy groups, are protected in order to avoidtheir unwanted participation in reactions. Protecting groups are used toblock some or all of the reactive moieties and prevent such groups fromparticipating in chemical reactions until the protective group isremoved. In other embodiments, each protective group is removable by adifferent means. Protective groups that are cleaved under totallydisparate reaction conditions fulfill the requirement of differentialremoval.

In certain embodiments, protective groups are removed by acid, base,reducing conditions (such as, for example, hydrogenolysis), and/oroxidative conditions. Groups such as trityl, dimethoxytrityl, acetal andt-butyldimethylsilyl are acid labile and are used to protect carboxy andhydroxy reactive moieties in the presence of amino groups protected withCbz groups, which are removable by hydrogenolysis, and Fmoc groups,which are base labile. Carboxylic acid and hydroxy reactive moieties areblocked with base labile groups such as, but not limited to, methyl,ethyl, and acetyl, in the presence of amines that are blocked with acidlabile groups, such as t-butyl carbamate, or with carbamates that areboth acid and base stable but hydrolytically removable.

In certain embodiments, carboxylic acid and hydroxy reactive moietiesare blocked with hydrolytically removable protective groups such as thebenzyl group, while amine groups capable of hydrogen bonding with acidsare blocked with base labile groups such as Fmoc. Carboxylic acidreactive moieties are protected by conversion to simple ester compoundsas exemplified herein, which include conversion to alkyl esters, or areblocked with oxidatively-removable protective groups such as2,4-dimethoxybenzyl, while co-existing amino groups are blocked withfluoride labile silyl carbamates.

Allyl blocking groups are useful in the presence of acid- andbase-protecting groups since the former are stable and are subsequentlyremoved by metal or pi-acid catalysts. For example, an allyl-blockedcarboxylic acid is deprotected with a palladium-catalyzed reaction inthe presence of acid labile t-butyl carbamate or base-labile acetateamine protecting groups. Yet another form of protecting group is a resinto which a compound or intermediate is attached. As long as the residueis attached to the resin, that functional group is blocked and does notreact. Once released from the resin, the functional group is availableto react.

Typically blocking/protecting groups may be selected from:

Other protecting groups, plus a detailed description of techniquesapplicable to the creation of protecting groups and their removal aredescribed in Greene & Wuts, Protective Groups in Organic Synthesis, 3rdEd., John Wiley & Sons, New York, N.Y., 1999, and Kocienski, ProtectiveGroups, Thieme Verlag, New York, N.Y., 1994, which are incorporatedherein by reference for such disclosure.

Compositions

The invention includes a pharmaceutical composition comprising at leastone compound of the invention and at least one pharmaceuticallyacceptable carrier. In certain embodiments, the composition isformulated for an administration route such as oral or parenteral, forexample, transdermal, transmucosal (e.g., sublingual, lingual,(trans)buccal, (trans)urethral, vaginal (e.g., trans- andperivaginally), (intra)nasal and (trans)rectal), intravesical,intrapulmonary, intraduodenal, intragastrical, intrathecal,subcutaneous, intramuscular, intradermal, intra-arterial, intravenous,intrabronchial, inhalation, and topical administration.

Methods

The invention includes a method of treating or preventing a diseaseassociated with and/or caused by overexpression and/or uncontrolledactivation of a tyrosine kinase in a subject in need thereof. Theinvention further includes a method of treating or preventing a cancerassociated with and/or caused by an oncogenic tyrosine kinase in asubject in need thereof. In certain embodiments, the disease comprises acancer. In other embodiments, the tyrosine kinase is c-ABL and/orBCR-ABL. In yet other embodiments, the cancer is chronic myelogenousleukemia (CML).

Examples of cancers that can be treated or prevented by the presentinvention include but are not limited to: squamous cell cancer, lungcancer including small cell lung cancer, non-small cell lung cancer,vulval cancer, thyroid cancer, adenocarcinoma of the lung and squamouscarcinoma of the lung, cancer of the peritoneum, hepatocellular cancer,gastric or stomach cancer including gastrointestinal cancer, pancreaticcancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer,bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer,colorectal cancer, endometrial or uterine carcinoma, salivary glandcarcinoma, kidney or renal cancer, prostate cancer, hepatic carcinoma,anal carcinoma, penile carcinoma, and head and neck cancer. In certainembodiments, the cancer is at least one selected from the groupconsisting of ALL, T-lineage Acute lymphoblastic Leukemia (T-ALL),T-lineage lymphoblastic Lymphoma (T-LL), Peripheral T-cell lymphoma,Adult T-cell Leukemia, Pre-B ALL, Pre-B Lymphomas, Large B-cellLymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphia chromosomepositive ALL, Philadelphia chromosome positive CML, lymphoma, leukemia,multiple myeloma myeloproliferative diseases, large B cell lymphoma, andB cell Lymphoma.

The methods of the invention comprise administering to the subject atherapeutically effective amount of at least one compound of theinvention, which is optionally formulated in a pharmaceuticalcomposition. In certain embodiments, the method further comprisesadministering to the subject an additional therapeutic agent that treatsor prevents cancer.

In certain embodiments, administering the compound of the invention tothe subject allows for administering a lower dose of the additionaltherapeutic agent as compared to the dose of the additional therapeuticagent alone that is required to achieve similar results in treating orpreventing a cancer in the subject. For example, in certain embodiments,the compound of the invention enhances the anti-cancer activity of theadditional therapeutic compound, thereby allowing for a lower dose ofthe additional therapeutic compound to provide the same effect.

In certain embodiments, the compound of the invention and thetherapeutic agent are co-administered to the subject. In otherembodiments, the compound of the invention and the therapeutic agent arecoformulated and co-administered to the subject.

In certain embodiments, the subject is a mammal. In other embodiments,the mammal is a human.

Combination Therapies

The compounds useful within the methods of the invention may be used incombination with one or more additional therapeutic agents useful fortreating a cancer. These additional therapeutic agents may comprisecompounds that are commercially available or synthetically accessible tothose skilled in the art. These additional therapeutic agents are knownto treat, prevent, or reduce the symptoms, of a cancer.

In non-limiting examples, the compounds useful within the invention maybe used in combination with one or more of the following therapeuticagents: Erlotinib (TARCEVA®, Genentech/OSI Pharm.), docetaxel(TAXOTERE®, Sanofi-Aventis), 5-FU (fluorouracil, 5-fluorouracil, CAS No.51-21-8), gemcitabine (GEMZAR®, Lilly), PD-0325901 (CAS No. 391210-10-9,Pfizer), cisplatin (cis-diamine, dichloroplatinum (II), CAS No.15663-27-1), carboplatin (CAS No. 41575-94-4), paclitaxel (TAXOL®,Bristol-Myers Squibb Oncology, Princeton, N.J.), pemetrexed (ALIMTA, EliLilly), trastuzumab (HERCEPTIN®, Genentech), temozolomide(4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo[4.3.0]nona-2,7,9-triene-9-carboxamide,CAS No. 85622-93-1, TEMODAR®, TEMODAL®, Schering Plough), tamoxifen((Z)-2-[4-(1,2-diphenylbut-1-enyl)phenoxy]-N,N-dimethylethanamine,NOLVADEX®, ISTUBAL®, VALODEX®), and doxorubicin (ADRIAMYCIN®), Akti-1/2,HPPD, rapamycin, oxaliplatin (ELOXATIN®, Sanofi), bortezomib (VELCADE®,Millennium Pharm.), sutent (SUNITINIB®, SU11248, Pfizer), letrozole(FEMARA®, Novartis), imatinib mesylate (GLEEVEC®, Novartis), XL-518 (Mekinhibitor, Exelixis, WO 2007/044515), ARRY-886 (Mek inhibitor, AZD6244,Array BioPharma, Astra Zeneca), SF-1126 (PI3K inhibitor, SemaforePharmaceuticals), BEZ-235 (PI3K inhibitor, Novartis), XL-147 (PI3Kinhibitor, Exelixis), PTK787/ZK 222584 (Novartis), fulvestrant(FASLODEX®, AstraZeneca), leucovorin (folinic acid), rapamycin(sirolimus, RAPAMUNE, Wyeth), lapatinib (TYKERB®, GSK572016, Glaxo SmithKline), lonafarnib (SARASAR™, SCH 66336, Schering Plough), sorafenib(NEXAVAR®, BAY43-9006, Bayer Labs), gefitinib (IRESSA®, AstraZeneca),irinotecan (CAMPTOSAR®, CPT-11, Pfizer), tipifarnib (ZARNESTRA™, Johnson& Johnson), ABRAXANE™ (Cremophor-free), albumin-engineered nanoparticleformulations of paclitaxel (American Pharmaceutical Partners,Schaumberg, Ill.), vandetanib (rINN, ZD6474, ZACTIMA®, AstraZeneca),chloranmbucil, AG1478, AG1571 (SU 5271; Sugen), temsirolimus (TORISEL®,Wyeth), pazopanib (GlaxoSmithKline), canfosfamide (TELCYTA®, Telik),thiotepa and cyclosphosphamide (CYTOXAN, NEOSAR®), ALK TKI inhibitors,antibodies such as avastin and cetuximab that target VEGFR and EGFRrespectively, other RTK TKIs for PDGFR or RET, immunotherapies such asipiliumimab and nivolumab, and radiation therapy.

In certain embodiments, the compounds of the present invention are usedin combination with radiation therapy. In other embodiments, thecombination of administration of the compounds of the present inventionand application of radiation therapy is more effective in treating orpreventing cancer than application of radiation therapy by itself. Inyet other embodiments, the combination of administration of thecompounds of the present invention and application of radiation therapyallows for use of lower amount of radiation therapy in treating thesubject.

A synergistic effect may be calculated, for example, using suitablemethods such as, for example, the Sigmoid-E_(max) equation (Holford &Scheiner, 1981, Clin. Pharmacokinet. 6:429-453), the equation of Loeweadditivity (Loewe & Muischnek, 1926, Arch. Exp. Pathol Pharmacol.114:313-326) and the median-effect equation (Chou & Talalay, 1984, Adv.Enzyme Regul. 22:27-55). Each equation referred to above may be appliedto experimental data to generate a corresponding graph to aid inassessing the effects of the drug combination. The corresponding graphsassociated with the equations referred to above are theconcentration-effect curve, isobologram curve and combination indexcurve, respectively.

Administration/Dosage/Formulations

The regimen of administration may affect what constitutes an effectiveamount. The therapeutic formulations may be administered to the subjecteither prior to or after the onset of a cancer. Further, several divideddosages, as well as staggered dosages may be administered daily orsequentially, or the dose may be continuously infused, or may be a bolusinjection. Further, the dosages of the therapeutic formulations may beproportionally increased or decreased as indicated by the exigencies ofthe therapeutic or prophylactic situation.

Administration of the compositions of the present invention to apatient, preferably a mammal, more preferably a human, may be carriedout using known procedures, at dosages and for periods of time effectiveto treat a cancer in the patient. An effective amount of the therapeuticcompound necessary to achieve a therapeutic effect may vary according tofactors such as the state of the disease or disorder in the patient; theage, sex, and weight of the patient; and the ability of the therapeuticcompound to treat a cancer in the patient. Dosage regimens may beadjusted to provide the optimum therapeutic response. For example,several divided doses may be administered daily or the dose may beproportionally reduced as indicated by the exigencies of the therapeuticsituation. A non-limiting example of an effective dose range for atherapeutic compound of the invention is from about 1 and 5,000 mg/kg ofbody weight/per day. One of ordinary skill in the art would be able tostudy the relevant factors and make the determination regarding theeffective amount of the therapeutic compound without undueexperimentation.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In particular, the selected dosage level depends upon a variety offactors including the activity of the particular compound employed, thetime of administration, the rate of excretion of the compound, theduration of the treatment, other drugs, compounds or materials used incombination with the compound, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well, known in the medical arts.

A medical doctor, e.g., physician or veterinarian, having ordinary skillin the art may readily determine and prescribe the effective amount ofthe pharmaceutical composition required. For example, the physician orveterinarian could start doses of the compounds of the inventionemployed in the pharmaceutical composition at levels lower than thatrequired in order to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulatethe compound in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the patients tobe treated; each unit containing a predetermined quantity of therapeuticcompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical vehicle. The dosage unitforms of the invention are dictated by and directly dependent on (a) theunique characteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding/formulating such a therapeutic compound for thetreatment of a cancer in a patient.

In certain embodiments, the compositions of the invention are formulatedusing one or more pharmaceutically acceptable excipients or carriers. Incertain embodiments, the pharmaceutical compositions of the inventioncomprise a therapeutically effective amount of a compound of theinvention and a pharmaceutically acceptable carrier.

The carrier may be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetable oils. The proper fluidity may be maintained, forexample, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms may be achievedby various antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it is preferable to include isotonic agents, for example, sugars,sodium chloride, or polyalcohols such as mannitol and sorbitol, in thecomposition. Prolonged absorption of the injectable compositions may bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate or gelatin.

In certain embodiments, the compositions of the invention areadministered to the patient in dosages that range from one to five timesper day or more. In other embodiments, the compositions of the inventionare administered to the patient in range of dosages that include, butare not limited to, once every day, every two, days, every three days toonce a week, and once every two weeks. It is readily apparent to oneskilled in the art that the frequency of administration of the variouscombination compositions of the invention varies from individual toindividual depending on many factors including, but not limited to, age,disease or disorder to be treated, gender, overall health, and otherfactors. Thus, the invention should not be construed to be limited toany particular dosage regime and the precise dosage and composition tobe administered to any patient is determined by the attending physicaltaking all other factors about the patient into account.

Compounds of the invention for administration may be in the range offrom about 1 μg to about 10,000 mg, about 20 μg to about 9,500 mg, about40 μg to about 9,000 mg, about 75 μg to about 8,500 mg, about 150 μg toabout 7,500 mg, about 200 μg to about 7,000 mg, about 350 μg to about6,000 mg, about 500 μg to about 5,000 mg, about 750 μg to about 4,000mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 30 mg toabout 1,000 mg, about 40 mg to about 900 mg, about 50 mg to about 800mg, about 60 mg to about 750 mg, about 70 mg to about 600 mg, about 80mg to about 500 mg, and any and all whole or partial incrementstherebetween.

In some embodiments, the dose of a compound of the invention is fromabout 1 mg and about 2,500 mg. In some embodiments, a dose of a compoundof the invention used in compositions described herein is less thanabout 10,000 mg, or less than about 8,000 mg, or less than about 6,000mg, or less than about 5,000 mg, or less than about 3,000 mg, or lessthan about 2,000 mg, or less than about 1,000 mg, or less than about 500mg, or less than about 200 mg, or less than about 50 mg. Similarly, insome embodiments, a dose of a second compound as described herein isless than about 1,000 mg, or less than about 800 mg, or less than about600 mg, or less than about 500 mg, or less than about 400 mg, or lessthan about 300 mg, or less than about 200 mg, or less than about 100 mg,or less than about 50 mg, or less than about 40 mg, or less than about30 mg, or less than about 25 mg, or less than about 20 mg, or less thanabout 15 mg, or less than about 10 mg, or less than about 5 mg, or lessthan about 2 mg, or less than about 1 mg, or less than about 0.5 mg, andany and all whole or partial increments thereof.

In certain embodiments, the present invention is directed to a packagedpharmaceutical composition comprising a container holding atherapeutically effective amount of a compound of the invention, aloneor in combination with a second pharmaceutical agent; and instructionsfor using the compound to treat, prevent, or reduce one or more symptomsof a cancer in a patient.

Formulations may be employed in admixtures with conventional excipients,i.e., pharmaceutically acceptable organic or inorganic carriersubstances suitable for oral, parenteral, nasal, intravenous,subcutaneous, enteral, or any other suitable mode of administration,known to the art. The pharmaceutical preparations may be sterilized andif desired mixed with auxiliary agents, e.g., lubricants, preservatives,stabilizers, wetting agents, emulsifiers, salts for influencing osmoticpressure buffers, coloring, flavoring and/or aromatic substances and thelike. They may also be combined where desired with other active agents,e.g., other analgesic agents.

Routes of administration of any of the compositions of the inventioninclude oral, nasal, rectal, intravaginal, parenteral, buccal,sublingual or topical. The compounds for use in the invention may beformulated for administration by any suitable route, such as for oral orparenteral, for example, transdermal, transmucosal (e.g., sublingual,lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- andperivaginally), (intra)nasal and (trans)rectal), intravesical,intrapulmonary, intraduodenal, intragastrical, intrathecal,subcutaneous, intramuscular, intradermal, intra-arterial, intravenous,intrabronchial, inhalation, and topical administration.

Suitable compositions and dosage forms include, for example, tablets,capsules, caplets, pills, gel caps, troches, dispersions, suspensions,solutions, syrups, granules, beads, transdermal patches, gels, powders,pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs,suppositories, liquid sprays for nasal or oral administration, drypowder or aerosolized formulations for inhalation, compositions andformulations for intravesical administration and the like. It should beunderstood that the formulations and compositions that would be usefulin the present invention are not limited to the particular formulationsand compositions that are described herein.

Oral Administration

For oral application, particularly suitable are tablets, dragees,liquids, drops, suppositories, or capsules, caplets and gelcaps. Thecompositions intended for oral use may be prepared according to anymethod known in the art and such compositions may contain one or moreagents selected from the group consisting of inert, non-toxicpharmaceutically excipients that are suitable for the manufacture oftablets. Such excipients include, for example an inert diluent such aslactose; granulating and disintegrating agents such as cornstarch;binding agents such as starch; and lubricating agents such as magnesiumstearate. The tablets may be uncoated or they may be coated by knowntechniques for elegance or to delay the release of the activeingredients. Formulations for oral use may also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertdiluent.

For oral administration, the compounds of the invention may be in theform of tablets or capsules prepared by conventional means withpharmaceutically acceptable excipients such as binding agents (e.g.,polyvinylpyrrolidone, hydroxypropylcellulose or hydroxypropylmethylcellulose); fillers (e.g., cornstarch, lactose, microcrystallinecellulose or calcium phosphate); lubricants (e.g., magnesium stearate,talc, or silica); disintegrates (e.g., sodium starch glycollate); orwetting agents (e.g., sodium lauryl sulphate). If desired, the tabletsmay be coated using suitable methods and coating materials such asOPADRY™ film coatingsystems avaiable from Colorcon, West Point, Pa.(e.g., OPADRY™ OY Type, OYC Type, Organic Enteric OY-P Type, AqueousEnteric OY-A Type, OY-PM Type and OPADRY™ White, 32K18400). Liquidpreparation for oral administration may be in the form of solutions,syrups or suspensions. The liquid preparations may be prepared byconventional means with pharmaceutically acceptable additives such assuspending agents (e.g., sorbitol syrup, methyl cellulose orhydrogenated edible fats); emulsifying agent (e.g., lecithin or acacia);non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol);and preservatives (e.g., methyl or propyl p-hydroxy benzoates or sorbicacid).

Granulating techniques are well known in the pharmaceutical art formodifying starting powders or other particulate materials of an activeingredient. The powders are typically mixed with a binder material intolarger permanent free-flowing agglomerates or granules referred to as a“granulation.” For example, solvent-using “wet” granulation processesare generally characterized in that the powders are combined with abinder material and moistened with water or an organic solvent underconditions resulting in the formation of a wet granulated mass fromwhich the solvent must then be evaporated.

Melt granulation generally consists in the use of materials that aresolid or semi-solid at room temperature (i.e. having a relatively lowsoftening or melting point range) to promote granulation of powdered orother materials, essentially in the absence of added water or otherliquid solvents. The low melting solids, when heated to a temperature inthe melting point range, liquefy to act as a binder or granulatingmedium. The liquefied solid spreads itself over the surface of powderedmaterials with which it is contacted, and on cooling, forms a solidgranulated mass in which the initial materials are bound together. Theresulting melt granulation may then be provided to a tablet press or beencapsulated for preparing the oral dosage form. Melt granulationimproves the dissolution rate and bioavailability of an active (i.e.drug) by forming a solid dispersion or solid solution.

U.S. Pat. No. 5,169,645 discloses directly compressible wax-containinggranules having improved flow properties. The granules are obtained whenwaxes are admixed in the melt with certain flow improving additives,followed by cooling and granulation of the admixture. In certainembodiments, only the wax itself melts in the melt combination of thewax(es) and additives(s), and in other cases both the wax(es) and theadditives(s) melt.

The present invention also includes a multi-layer tablet comprising alayer providing for the delayed release of one or more compounds of theinvention, and a further layer providing for the immediate release of amedication for treatment of G-protein receptor-related diseases ordisorders. Using a wax/pH-sensitive polymer mix, a gastric insolublecomposition may be obtained in which the active ingredient is entrapped,ensuring its delayed release.

Parenteral Administration

For parenteral administration, the compounds of the invention may beformulated for injection or infusion, for example, intravenous,intramuscular or subcutaneous injection or infusion, or foradministration in a bolus dose and/or continuous infusion. Suspensions,solutions or emulsions in an oily or aqueous vehicle, optionallycontaining other formulatory agents such as suspending, stabilizingand/or dispersing agents may be used.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1, 3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. Sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose, any bland fixed oil may beemployed including synthetic mono- or di-glycerides. Fatty acids, suchas oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such as Ph. Helv orsimilar alcohol.

Additional Administration Forms

Additional dosage forms of this invention include dosage forms asdescribed in U.S. Pat. Nos. 6,340,475; 6,488,962; 6,451,808; 5,972,389;5,582,837; and 5,007,790. Additional dosage forms of this invention alsoinclude dosage forms as described in U.S. Patent Applications Nos.20030147952; 20030104062; 20030104053; 20030044466; 20030039688; and20020051820. Additional dosage forms of this invention also includedosage forms as described in PCT Applications Nos. WO 03/35041; WO03/35040; WO 03/35029; WO 03/35177; WO 03/35039; WO 02/96404; WO02/32416; WO 01/97783; WO 01/56544; WO 01/32217; WO 98/55107; WO98/11879; WO 97/47285; WO 93/18755; and WO 90/11757.

Controlled Release Formulations and Drug Delivery Systems

In certain embodiments, the formulations of the present invention maybe, but are not limited to, short-term, rapid-offset, as well ascontrolled, for example, sustained release, delayed release andpulsatile release formulations.

The term sustained release is used in its conventional sense to refer toa drug formulation that provides for gradual release of a drug over anextended period of time, and that may, although not necessarily, resultin substantially constant blood levels of a drug over an extended timeperiod. The period of time may be as long as a month or more and shouldbe a release which is longer that the same amount of agent administeredin bolus form.

For sustained release, the compounds may be formulated with a suitablepolymer or hydrophobic material which provides sustained releaseproperties to the compounds. As such, the compounds for use the methodof the invention may be administered in the form of microparticles, forexample, by injection or in the form of wafers or discs by implantation.

In one embodiment of the invention, the compounds of the invention areadministered to a patient, alone or in combination with anotherpharmaceutical agent, using a sustained release formulation.

The term delayed release is used herein in its conventional sense torefer to a drug formulation that provides for an initial release of thedrug after some delay following drug administration and that mat,although not necessarily, includes a delay of from about 10 minutes upto about 12 hours.

The term pulsatile release is used herein in its conventional sense torefer to a drug formulation that provides release of the drug in such away as to produce pulsed plasma profiles of the drug after drugadministration.

The term immediate release is used in its conventional sense to refer toa drug formulation that provides for release of the drug immediatelyafter drug administration.

As used herein, short-term refers to any period of time up to andincluding about 8 hours, about 7 hours, about 6 hours, about 5 hours,about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40minutes, about 20 minutes, or about 10 minutes and any or all whole orpartial increments thereof after drug administration after drugadministration.

As used herein, rapid-offset refers to any period of time up to andincluding about 8 hours, about 7 hours, about 6 hours, about 5 hours,about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40minutes, about 20 minutes, or about 10 minutes, and any and all whole orpartial increments thereof after drug administration.

Dosing

The therapeutically effective amount or dose of a compound of thepresent invention depends on the age, sex and weight of the patient, thecurrent medical condition of the patient and the progression of a cancerin the patient being treated. The skilled artisan is able to determineappropriate dosages depending on these and other factors.

A suitable dose of a compound of the present invention may be in therange of from about 0.01 mg to about 5,000 mg per day, such as fromabout 0.1 mg to about 1,000 mg, for example, from about 1 mg to about500 mg, such as about 5 mg to about 250 mg per day. The dose may beadministered in a single dosage or in multiple dosages, for example from1 to 4 or more times per day. When multiple dosages are used, the amountof each dosage may be the same or different. For example, a dose of 1 mgper day may be administered as two 0.5 mg doses, with about a 12-hourinterval between doses.

It is understood that the amount of compound dosed per day may beadministered, in non-limiting examples, every day, every other day,every 2 days, every 3 days, every 4 days, or every 5 days. For example,with every other day administration, a 5 mg per day dose may beinitiated on Monday with a first subsequent 5 mg per day doseadministered on Wednesday, a second subsequent 5 mg per day doseadministered on Friday, and so on.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion the administration of the inhibitor of the invention isoptionally given continuously; alternatively, the dose of drug beingadministered is temporarily reduced or temporarily suspended for acertain length of time (i.e., a “drug holiday”). The length of the drugholiday optionally varies between 2 days and 1 year, including by way ofexample only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days,12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days,120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days,320 days, 350 days, or 365 days. The dose reduction during a drugholiday includes from 10%-100%, including, by way of example only, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, or 100%.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, is reduced to a level at which theimproved disease is retained. In certain embodiments, patients requireintermittent treatment on a long-term basis upon any recurrence ofsymptoms and/or infection.

The compounds for use in the method of the invention may be formulatedin unit dosage form. The term “unit dosage form” refers to physicallydiscrete units suitable as unitary dosage for patients undergoingtreatment, with each unit containing a predetermined quantity of activematerial calculated to produce the desired therapeutic effect,optionally in association with a suitable pharmaceutical carrier. Theunit dosage form may be for a single daily dose or one of multiple dailydoses (e.g., about 1 to 4 or more times per day). When multiple dailydoses are used, the unit dosage form may be the same or different foreach dose.

Toxicity and therapeutic efficacy of such therapeutic regimens areoptionally determined in cell cultures or experimental animals,including, but not limited to, the determination of the LD₅₀ (the doselethal to 50% of the population) and the ED₅₀ (the dose therapeuticallyeffective in 50% of the population). The dose ratio between the toxicand therapeutic effects is the therapeutic index, which is expressed asthe ratio between LD₅₀ and ED₅₀. The data obtained from cell cultureassays and animal studies are optionally used in formulating a range ofdosage for use in human. The dosage of such compounds lies preferablywithin a range of circulating concentrations that include the ED₅₀ withminimal toxicity. The dosage optionally varies within this rangedepending upon the dosage form employed and the route of administrationutilized.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures, embodiments, claims, and examples described herein.Such equivalents were considered to be within the scope of thisinvention and covered by the claims appended hereto. For example, itshould be understood, that modifications in reaction conditions,including but not limited to reaction times, reaction size/volume, andexperimental reagents, such as solvents, catalysts, pressures,atmospheric conditions, e.g., nitrogen atmosphere, andreducing/oxidizing agents, with art-recognized alternatives and using nomore than routine experimentation, are within the scope of the presentapplication.

It is to be understood that wherever values and ranges are providedherein, all values and ranges encompassed by these values and ranges,are meant to be encompassed within the scope of the present invention.Moreover, all values that fall within these ranges, as well as the upperor lower limits of a range of values, are also contemplated by thepresent application.

The following examples further illustrate aspects of the presentinvention. However, they are in no way a limitation of the teachings ordisclosure of the present invention as set forth herein.

Examples

The invention is now described with reference to the following Examples.These Examples are provided for the purpose of illustration only and theinvention should in no way be construed as being limited to theseExamples, but rather should be construed to encompass any and allvariations which become evident as a result of the teaching providedherein.

Methods and Materials 1. Biology Cell Lines and Materials

K562 cells were obtained from ATCC and were grown at 37° C., 5% CO₂ inIscove's Modified Dulbecco's Media (IMDM) supplemented with 10% FBS, 100U/mL penicillin and 100 μg/mL streptomycin. HEK293 and SK-BR-3 cellswere obtained from ATCC and were grown at 37° C., 5% CO₂ in Dulbecco'sModified Eagle's Media (DMEM) supplemented with 10% FBS, 100 U/mLpenicillin and 100 μg/mL streptomycin. Phospho-STAT5 Y694 (#4322) andphospho-CrkL Y207 (#3181) antibodies were obtained from Cell SignalingTechnologies. c-ABL (24-11) antibody was obtained from Santa CruzBiotechnologies. α-Tubulin antibody (T9026) was purchased fromSigma-Aldrich.

Western Blotting

K562 cells (1-1.5×10⁶) were treated for 24 hours with the indicatedcompounds solubilized in DMSO. The cells were collected at 300 g for 3min. The cells were then lysed in lysis buffer (25 mM Tris, 1% Triton,0.25% deoxycholic acid) with Roche protease inhibitor complete cocktailand phosphatase inhibitors (10 mM sodium fluoride, 10 mM sodiumpyrophosphate, 1 mM sodium orthovanadate and 20 mM 0-glycerophosphate).The total protein concentrations were determined by Pierce BCA ProteinAssay and 30-50 g of protein was loaded onto 10% Tris-Glycine gels.After standard gel electrophoresis, the separated proteins weretransferred to nitrocellulose by wet transfer. The immunoblots were thenprocessed by standard procedures and incubated with the respectiveantibodies. Band intensities were quantified by Bio-Rad's Image Labsoftware.

In Vitro Kinase Binding Affinity Determination

PROTAC in vitro binding affinities (Kd) to phosphorylated andnon-phosphorylated c-ABL kinase domain were determined using theKinomeScan platform (DiscoverRx Corporation). The compounds weresolubilized in DMSO and sent to DiscoverRx Corporation as a 10 μM stocksolution.

Cell Viability Assay

HEK293T and SK-BR-3 were washed thrice with 1× phosphate-buffered saline(PBS), trypsinized and 6000 cells were plated in triplicates on a tissuecultured treated 96-well plate with 50 μL of DMEM. After 24 hours,PROTAC was added directly on top of cells in 50 μL of DMEM. After 48hours of PROTAC treatment, a CellTiter-Glo® Luminescent Cell Viabilityassay (Promega) was performed as detailed in the manufacturer's manual.The data was analyzed using nonlinear regression in GraphPad Prism® 6.Similarly, 6000 cells of K562 suspension cell line were plated in atissue cultured treated 96-well plate with 50 μL of IMIDM. PROTAC wasadded directly on top of cells in 50 μL of IMDM. After 48 hours ofPROTAC treatment, a CellTiter-Glo® Luminescent Cell Viability assay(Promega) was performed as detailed in the manufacturer's manual. TheY-axis of FIG. 4 corresponds to the luminescence of PROTAC-treatedsamples normalized to the luminescence of DMSO-treated samples.

2. Chemistry General Methods

All reactions were carried out under an atmosphere of dry nitrogen orargon. Glassware was oven-dried prior to use. Unless otherwiseindicated, common reagents or materials were obtained from commercialsource and used without further purification. N,N-Diisopropylethylamine(DIPEA) was obtained anhydrous by distillation over potassium hydroxide.Tetrahydrofuran (THF), Dichloromethane (CH₂Cl₂), and dimethylforamide(DMF) was dried by a PureSolvm solvent drying system. Flash columnchromatography was performed using silica gel 60 (230-400 mesh).Analytical thin layer chromatography (TLC) was carried out on Mercksilica gel plates with QF-254 indicator and visualized by UV or KMnO4.¹H and ¹³C NMR spectra were recorded on an Agilent DD₂ 500 (500 MHz H;125 MHz ¹³C) or Agilent DD₂ 600 (600 MHz H; 150 MHz ¹³C) or Agilent DD₂400 (400 MHz H; 100 MHz ¹³C) spectrometer at room temperature. Chemicalshifts were reported in ppm relative to the residual CDCl₃ (δ 7.26 ppmH; δ 77.0 ppm ¹³C), CD₃OD (δ 3.31 ppm H; δ 49.00 ppm ¹³C), or d⁶-DMSO (δ2.50 ppm H; δ 39.52 ppm ¹³C). NMR chemical shifts were expressed in ppmrelative to internal solvent peaks, and coupling constants were measuredin Hz. (bs=broad signal). In most cases, only peaks of the major rotamerare reported. Mass spectra were obtained using Agilent 1100 seriesLC/MSD spectrometers. Analytical HPLC analyses were carried out on250×4.6 mm C-18 column using gradient conditions (10-100% B, flowrate=1.0 mL/min, 20 min). Preparative HPLC was carried out on 250×21.2mm C-18 column using gradient conditions (10-100% B, flow rate=10.0mL/min, 20 min). The eluents used were: solvent A (H₂O with 0.1% TFA)and solvent B (CH₃CN with 0.1% TFA).

Synthesis of the Linkers

tert-Butyl 5-(2-(2-((6-chlorohexyl)oxy)ethoxy)ethoxy)pentanoate

To a solution of 2-(2-((6-chlorohexyl)oxy)ethoxy)ethan-1-ol (0.98 ml,7.32 mmol) and tert-butyl 6-bromohexanoate (1.84 g, 7.32 mmol) inbenzene (6 mL) was added aqueous 50% NaOH (4 ml, 50 mmol) and TBAHS(2.49 g, 7.32 mmol). The reaction mixture was stirred vigorously at roomtemperature for 12 h (overnight). Then the reaction was diluted withether (50 mL) and water (50 mL), organic layer was separated, washedwith water (2×20 mL), dried (Na₂SO₄) and evaporated under vacuum. Crudeproduct was purified by column flash chromatography (SiO₂-120 g,gradient; Hex 100% to Hex:AcOEt, 95:5) to give the desired product (55%yield) as an oil. ¹H NMR (400 MHz, Chloroform-d) δ 3.66-3.61 (m, 4H),3.57 (m, 4H), 3.52 (t, J=6.7 Hz, 2H), 3.45 (td, J=6.6, 2.1 Hz, 4H), 2.20(t, J=7.5 Hz, 2H), 1.77 (p, J=6.8 Hz, 2H), 1.59 (m, 6H), 1.43 (s, 9H),1.50-1.29 (m, 12H). ¹³C NMR (151 MHz, Chloroform-d) δ 173.28, 80.12,71.39, 71.35, 70.79, 70.27, 70.26, 45.21, 35.67, 32.71, 29.62, 29.48,28.27, 26.86, 25.76, 25.59, 25.08. HRMS (ESI); m/z: [M+Na]⁺ calcd forC₂₀H₃₉ClO₅Na: 417.2383, found 417.2376.

tert-Butyl 6-((5-((6-chlorohexyl)oxy)pentyl)oxy)hexanoate

Yield (29%); ¹H NMR (500 MHz, Chloroform-d) δ 3.52 (t, J=6.8, 2H),3.41-3.36 (m, 8H), 2.20 (t, J=7.8, 2H), 1.80-1.74 (m, 2H), 1.63-1.53 (m,10H), 1.45-1.34 (m, 17H). ¹³C NMR (100 MHz, Chloroform-d) δ 173.14,79.94, 70.81, 70.78, 70.68, 70.65, 45.05, 35.51, 32.55, 29.58, 29.43,28.09, 26.71, 25.69, 25.52, 24.94, 22.81. MS (ESI); m/z: [M+Na]⁺ Calcd.for C₂₁H₄₁CO₄Na, 415.2591. Found 415.2632.

Synthesis of tert-butyl 2-(2-((6-chlorohexyl)oxy)ethoxy)acetate

To a solution of 2-((6-chlorohexyl)oxy)ethan-1-ol (1.6 g, 8.85 mmol) ina mixture of DMF:THF (1:1), 20 mL was added NaH (95%, 268.5 mg, 10.6mmol) at 0° C., the reaction mixture was stirred for 30 min. at the sametemperature. tert-Butyl 2-bromoacetate (2.6 mL, 17.7 mmol) was thenadded at 0° C. and the reaction mixture was stirred for 12 h. Afterdilution with ethyl acetate (100 mL) and water (100 mL), the aqueousphase was extracted with ethyl acetate (3×100 mL). The combined organicfractions were washed with brine and dried over MgSO₄. Afterconcentration, the crude material was subjected to column chromatographyon silica gel (hexane/AcOEt 7/1) to give 1.67 g (64%) of tert-butyl2-(2-((6-chlorohexyl)oxy)ethoxy)acetate as a colorless oil. ¹H NMR (400MHz, Chloroform-d) δ 4.01 (s, 2H), 3.70-3.66 (m, 2H), 3.60-3.59 (m, 2H),3.54-3.49 (m, 2H), 3.48-3.43 (m, 2H), 1.80-1.73 (m, 2H), 1.61-155 (m,2H), 1.48-1.40 (m, 11H), 1.38-1.33 (m, 2H). ¹³C NMR (151 MHz,Chloroform-d) δ 169.64, 81.45, 71.23, 70.70, 70.11, 69.01, 45.00, 32.51,29.41, 28.07, 26.66, 25.39. HRMS (ESI); m/z: [M+Na]⁺ Calcd. forC₁₄H₂₇ClO₄Na, 317.1496. Found 317.1536.

tert-Butyl 24-chloro-3,6,9,12,15,18-hexaoxatetracosanoate

Yield (68%); ¹H NMR (500 MHz, Chloroform-d) δ 3.99 (s, 2H), 3.70-3.61(m, 18H), 3.57-3.53 (m, 2H), 3.50 (t, J=6.8 Hz, 2H), 3.42 (t, J=6.8 Hz,2H), 1.74 (p, J=7.3 Hz, 2H), 1.56 (p, J=7.3 Hz, 2H), 1.47-1.39 (m, 11H),1.36-1.31 (m, 2H). ¹³C NMR (151 MHz, Chloroform-d) δ 169.64, 81.48,71.17, 70.67, 70.56, 70.55, 70.53, 70.51, 70.05, 68.98, 45.05, 32.50,29.41, 28.06, 26.65, 25.37. HRMS (ESI): m/z; [M+Na]⁺ Calcd. forC₂₂H₄₃ClO₈Na: 493.2544 Found: 493.2649.

Synthesis of Dasatinib and Imatinib with VHL Ligand

(2S,4R)-1-((S)-2-(tert-butyl)-22-chloro-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

To a solution of 6-[2-[2-(6-chlorohexoxy)ethoxy]ethoxy]hexanoic acid (80mg, 0.24 mmol) in DMF (5 mL) was added HATU (179.53 mg, 0.47 mmol) andthe resulting solution was stirred for 10 minutes at rt, after which((2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide;hydrochloride (110.25 mg, 0.24 mmol) and DIEA(0.2 ml, 1.18 mmol) were added respectively. The resulting mixture wasstirred at room temperature for 16 h at rt. The product was extractedtwice with Ac₂O then purified by by silica gel DCM/MeOH 95:5 to give 152mg (85.7%) of the desired product. ¹H NMR (500 MHz, CD₃OD) δ 8.86 (s,1H), 7.45 (d, J=7.8 Hz, 2H), 7.41 (d, J=7.8 Hz, 2H), 4.69 (bs, 1H),4.58-4.49 (m, 3H), 4.34 (d, J=15.0 Hz, 1H), 3.89 (d, J=11.0 Hz, 1H),3.80 (d, J=11.0 Hz, 1H), 3.61-3.53 (m, 10H), 3.47-3.45 (m, 4H), 2.46 (s,3H), 2.32-2.19 (m, 3H), 2.11-2.06 (m, 1H), 1.78-1.72 (m, 2H), 1.65-1.54(m, 6H), 1.47-1.35 (m, 6H), 1.03 (s, 9H). ¹³C NMR (151 MHz, CD₃OD) δ174.4, 173.0, 170.9, 151.5, 147.5, 138.9, 132.0, 130.0, 128.9, 127.6,70.74, 70.71, 70.2, 69.8, 69.7, 59.4, 59.3, 57.5, 56.6, 44.3, 42.4,42.3, 37.5, 35.2, 35.1, 32.4, 29.2, 29.0, 26.3, 25.7, 25.5, 25.4, 25.1,27.3, 15.9, 14.5. MS (ESI); m/z: [M+H]⁺ calcd for C₃₈H₆₀ClN₄O₇S: 751.4,found 751.1.

(2S,4R)-1-((S)-2-(tert-butyl)-22-iodo-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

To a solution of (2S,4R)-1-[(2S)-2-[6-[2-[2-(6-chlorohexoxy)ethoxy]ethoxy]hexanoylamino]-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide(87 mg, 0.12 mmol) in acetone (10 ml) was added NaI (86.77 mg, 0.58mmol). The reaction mixture was stirred at reflux temperature for 24 h,then the solvent was removed under vacuum and crude product wasdissolved in EtOAc (15 mL) and an aqueous solution of Na₂SO₃ (10%, 10mL), organic layer was separated, washed with water (10 mL), dried(Na₂SO₄) and evaporated under vacuum. Crude product was pure by NMR(>98% purity), 96 mg (99%) of the desired product. It was used in thenext step without any further purification. ¹H NMR (500 MHz, CD₃OD) δ8.87 (s, 1H), 7.45 (d, J=7.8 Hz, 2H), 7.41 (d, J=7.8 Hz, 2H), 4.65-4.62(m, 1H), 4.58-4.49 (m, 3H), 4.37-4.32 (m, 1H), 3.89 (d, J=11.0 Hz, 1H),3.80 (d, J=11.0 Hz, 1H), 3.62-3.58 (m, 4H), 3.57-3.54 (m, 4H), 3.46 (m,4H), 3.26 (t, J=6.6 Hz, 2H), 2.47 (s, 3H), 2.32-2.19 (m, 3H), 2.11-2.06(m, 1H), 1.83-1.72 (m, 2H), 1.64-1.54 (m, 6H), 1.44-1.38 (m, 6H), 1.03(s, 9H). MS (ESI); m/z: [M+H]⁺ calcd for C₃₈H₆₀IN₄O_(7S): 843.3, found843.1.

N-(2-chloro-6-methylphenyl)-2-((6-(4-((S)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-11,14,17-trioxa-4-azatricosan-23-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-2-2-6-VHL

To a solution ofN-(2-chloro-6-methyl-phenyl)-2-[(2-methyl-6-piperazin-1-yl-pyrimidin-4-yl)amino]thiazole-5-carboxamide;2,2,2-trifluoroacetaldehyde(9.65 mg, 0.02 mmol) and DIEA (5.38 mg, 0.05 mmol) in DMF (1 ml) wasadded(2S,4R)-4-hydroxy-1-[(2S)-2-[6-[2-[2-(6-iodohexoxy)ethoxy]ethoxy]hexanoylamino]-3,3-dimethyl-butanoyl]-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide(15 mg, 0.02 mmol) and the resulting solution stirred for 16 h at rt.The solvent was evaporated and the residue subjected to Prep TLCpurification (ammonia/MeOH/DCM:1/10/60) to give 10.5 mg (50.9%) of thedesired product as a foamy brown solid. ¹H NMR (500 MHz, CD₃OD) δ 8.86(s, 1H), 8.14 (s, 1H), 7.45 (d, J=7.8 Hz, 2H), 7.41 (d, J=7.8 Hz, 2H),7.35 (d, J=7.6 Hz, 1H), 7.26-7.21 (m, 2H), 6.00 (s, 1H), 4.63 (bs, 1H),4.58-4.49 (m, 3H), 4.36 (d, J=15.0 Hz, 1H), 3.90 (d, J=11.0 Hz, 1H),3.80 (dd, J=11.0 Hz, J=3.8 Hz, 1H), 3.64-3.59 (m, 8H), 3.58-3.55 (m,4H), 3.49-3.45 (m, 4H), 2.55 (bs, 4H), 2.46 (s, 6H), 2.44-2.39 (m, 2H),2.32 (s, 3H), 2.29-2.19 (m, 3H), 2.10-2.06 (m, 1H), 1.64-1.55 (m, 8H),1.42-1.34 (m, 6H), 1.03 (s, 9H). ¹³C NMR (151 MHz, CD₃OD) δ 173.9,171.9, 171.8, 170.9, 166.1, 163.7, 162.8, 161.1, 156.6, 150.7, 147.6,140.3, 138.5, 138.1, 132.2, 132.0, 131.8, 130.1, 129.1, 128.8, 127.9,127.6, 126.8, 125.3, 83.8, 71.1, 70.9, 70.19, 70.18, 69.9, 69.5, 58.96,58.92, 58.4, 57.2, 56.7, 52.4, 49.4, 43.4, 42.6, 36.6, 36.8, 35.3, 29.2,28.9, 27.1, 26.1, 25.9, 25.7, 25.6, 25.4, 25.2, 25.0, 18.2, 15.2. MS(ESI); m/z: [M+H] calcd for C₅₈H₈₁ClN₁₁O₈S₂: 1158.5, found 1158.4.

(2S,4R)-1-((S)-2-(tert-butyl)-22-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:IMA-6-2-2-6-VHL

Yield (61%); ¹H NMR (500 MHz, CDCl₃) δ 9.22 (d, J=2.0 Hz, 1H), 8.69-8.66(m, 2H), 8.54 (s, 1H), 8.49-8.47 (m, 2H), 8.19 (s, 1H), 7.83 (d, J=8.1Hz, 2H), 7.42-7.29 (m, 9H), 7.19-7.15 (m, 3H), 6.25 (d, J=9.0 Hz, 1H),4.69 (t, J=7.9 Hz, 1H), 4.57-4.49 (m, 3H), 4.31 (d, J=14.9 Hz, J=5.1 Hz,1H), 4.01 (d, J=11.2 Hz, 1H), 3.63-3.53 (m, 10H), 3.47-3.39 (m, 7H),2.55-2.44 (m, 10H), 2.37-2.34 (m, 2H), 2.32 (s, 3H), 2.20-2.08 (m, 4H),1.61-1.44 (m, 8H), 1.36-1.34 (m, 6H), 0.91 (s, 9H). ¹³C NMR (151 MHz,CDCl₃) δ 173.5, 171.8, 170.8, 165.3, 162.7, 160.6, 158.9, 151.4, 150.3,148.43, 148.40, 142.2, 138.0, 137.7, 136.6, 134.9, 133.9, 132.7, 131.6,130.9, 130.7, 129.4, 129.3, 128.0, 127.1, 124.7, 123.7, 115.6, 113.7,108.2, 71.3, 71.1, 70.6, 70.5, 70.0, 69.9, 62.5, 58.6, 57.3, 56.7, 53.1,52.8, 50.7, 46.3, 43.2, 36.3, 35.9, 35.1, 29.5, 29.2, 27.3, 26.5, 26.4,25.9, 25.7, 25.3, 17.7, 16.0. MS (ESI); m/z: [M+H] calcd forC₆₆H₈₈N₁₁O₈S: 1194.6, found 1194.9.

(2S,4R)-1-((S)-2-(2-(2-((6-chlorohexyl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Yield (86%); ¹H NMR (500 MHz, CDCl₃) δ 8.70 (s, 1H), 7.45 (s, 1H),7.40-7.32 (m, 5H), 4.76-4.70 (m, 1H), 4.56-4.51 (m, 2H), 4.37 (d, J=15.0Hz, 1H), 4.08-3.97 (m, 3H), 3.75-3.60 (m, 6H), 3.55-3.48 (m, 3H),3.23-3.20 (m, 2H), 2.53 (s, 3H), 2.52-2.48 (m, 1H), 2.16-2.21 (m, 1H),1.82-1.74 (m, 2H), 1.63-1.59 (m, 2H), 1.51-1.38 (m, 4H), 0.99 (s, 9H).¹³C NMR (151 MHz, CDCl₃) δ 171.2, 170.9, 170.5, 150.3, 148.4, 138.1,131.8, 130.8, 129.4, 128.0, 71.3, 71.2, 70.2, 70.1, 69.7, 58.6, 57.2,56.6, 55.7, 45.0, 43.6, 43.1, 36.0, 34.9, 32.5, 29.3, 26.6, 26.4, 25.3,16.0. MS (ESI); m/z: [M+H]⁺ calcd for C₃₂H₄₈ClN₄O₆S: 651.3, found 651.1.

(2S,4R)-4-hydroxy-1-((S)-2-(2-(2-((6-iodohexyl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Yield (89%); ¹H NMR (500 MHz, CD₃OD) δ 8.89 (s, 1H), 7.48-7.45 (m, 2H),7.43-7.41 (m, 2H), 4.68-4.66 (m, 1H), 4.60-4.55 (m, 2H), 4.37 (d, J=15.5Hz, 1H), 4.05 (s, 2H), 3.90 (d, J=11.0 Hz, 1H), 3.79 (s, 3H), 3.75-3.70(m, 2H), 3.65-3.61 (m, 2H), 3.53-3.48 (m, 2H), 3.19 (t, J=6.8 Hz, 1H),2.47 (s, 3H), 2.26-2.24 (m, 1H), 2.12-2.01 (m, 1H), 1.80-1.74 (m, 2H),1.61-1.58 (m, 2H), 1.41-1.38 (m, 4H), 1.04 (s, 9H). MS (ESI); m/z:[M+H]⁺ calcd for C₃₂H₄₈IN₄O₆S: 743.2, found 743.0.

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-2-2-VHL

Yield (27%); ¹H NMR (500 MHz, CD₃OD) δ 8.86 (s, 1H), 8.14 (s, 1H),7.47-7.39 (m, 4H), 7.34 (d, J=7.3 Hz, 1H), 7.27-7.20 (m, 2H), 6.00 (s,1H), 4.69 (bs, 1H), 4.59-4.56 (m, 1H), 4.52-4.51 (m, 1H), 4.35 (d,J=15.6 Hz, 1H), 4.04 (s, 2H), 3.88 (d, J=11.0 Hz, 1H), 3.80 (dd, J=15.0Hz, J=3.8 Hz, 1H), 3.76-3.60 (m, 8H), 3.55-3.49 (m, 2H), 3.21-3.17 (m,1H), 2.56 (bs, 4H), 2.48-2.40 (m, 6H), 2.32 (s, 3H), 2.25-2.21 (m, 1H),2.12-2.07 (m, 1H), 1.63-1.55 (m, 4H), 1.43-1.28 (m, 6H), 1.05 (s, 9H).¹³C NMR (151 MHz, CD₃OD) δ 172.9, 170.7, 170.2, 166.0, 163.8, 162.9,161.8, 157.2, 151.4, 147.6, 140.7, 138.9, 138.8, 132.9, 132.8, 131.9,130.0, 129.1, 128.9, 128.7, 128.1, 128.0, 127.5, 126.9, 125.4, 82.5,70.97, 70.91, 69.66, 69.63, 69.60, 59.4, 58.1, 56.7, 56.7, 52.3, 48.1,43.2, 42.3, 37.5, 35.6, 29.2, 26.9, 25.8, 25.7, 25.6, 25.5, 24.2, 17.3,14.4. MS (ESI); m/z: [M+H]⁺ calcd for C₂H₆₉ClN₁₁O₇S₂: 1058.4, found1058.2.

(2S,4R)-1-((S)-3,3-dimethyl-2-(2-(2-((6-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:IMA-6-2-2-VHL

Yield (66%); ¹H NMR (500 MHz, CDCl₃/d₆DMSO) δ 9.23 (d, J=2.0 Hz, 1H),8.69-8.66 (m, 2H), 8.57 (s, 1H), 8.51-8.48 (m, 2H), 8.23 (s, 1H), 7.83(d, J=8.0 Hz, 2H), 7.47-7.38 (m, 4H), 7.35-7.27 (m, 6H), 7.20-7.16 (m,2H), 7.08 (s, 1H), 4.69 (t, J=7.9 Hz, 1H), 4.56-4.47 (m, 3H), 4.32 (d,J=15.1 Hz, J=5.3 Hz, 1H), 4.03-3.98 (m, 3H), 3.66-3.5 (m, 8H), 3.43 (t,J=6.5 Hz, 2H), 2.72-2.55 (m, 7H), 2.52-2.45 (m, 6H), 2.33 (s, 3H),2.14-2.08 (m, 1H), 1.58-1.50 (m, 4H), 1.36-1.25 (m, 6H), 0.94 (s, 9H).¹³C NMR (151 MHz, CDCl₃/d₆DMSO) δ 171.1, 170.9, 170.4, 165.6, 162.7,160.5, 159.0, 151.4, 150.3, 148.5, 148.4, 141.7, 138.1, 137.7, 136.6,134.9, 134.0, 132.7, 131.6, 130.8, 130.7, 129.4, 129.3, 128.0, 127.2,124.4, 123.8, 115.5, 113.4, 108.3, 71.3, 71.2, 70.3, 69.9, 69.8, 62.2,58.6, 58.2, 57.1, 56.7, 54.6, 52.8, 51.9, 50.7, 43.1, 36.1, 35.0, 29.2,27.0, 25.8, 17.7, 16.0. MS (ESI); m/z: [M+H] calcd for C₆₀H₇₆N₁₁O₇S:1094.5, found 1094.2.

(2S,4R)-1-((S)-2-(tert-butyl)-27-chloro-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Yield (77%); ¹H NMR (500 MHz, CDCl₃) δ 8.69 (s, 1H), 7.47-7.37 (m, 3H),7.30-7.29 (m, 3H), 4.74-4.69 (m, 1H), 4.58-4.52 (m, 2H), 4.40-4.35 (m,1H), 4.07-3.98 (m, 3H), 3.72-3.57 (m, 17H), 3.50-3.45 (m, 9H), 3.22-3.18(m, 1H), 2.55-2.46 (m, 4H), 2.18-2.14 (m, 1H), 1.82-1.76 (m, 2H),1.64-1.58 (m, 2H), 1.45-1.36 (m, 4H), 0.97 (s, 9H). NMR (151 MHz, CDCl₃)δ 171.1, 170.9, 170.4, 150.3, 148.3, 138.2, 131.6, 130.8, 129.4, 128.0,71.2, 71.0, 70.41, 70.38, 70.32, 70.27, 70.2, 69.98, 69.89, 58.6, 56.9,56.7, 55.2, 50.6, 45.0, 43.2, 43.1, 36.5, 35.2, 31.4, 29.3, 26.6, 26.3,25.3, 16.0. MS (ESI); m/z: [M+H]⁺ calcd for C₄₀H₆₄CN₄O₁₀S: 827.4, found827.2.

(2S,4R)-1-((S)-2-(tert-butyl)-27-iodo-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Yield (96%); ¹H NMR (500 MHz, CD₃OD) δ 8.88 (s, 1H), 7.47 (d, J=8.3 Hz,2H), 7.42 (d, J=8.3 Hz, 2H), 4.68 (bs, 1H), 4.58-4.51 (m, 3H), 4.36 (d,J=15.4 Hz, 1H), 4.07 (s, 2H), 3.87 (d, J=11.0 Hz, 1H), 3.81 (dd, J=11.0Hz, J=3.9 Hz, 1H), 3.75-3.61 (m, 18H), 3.58-3.55 (m, 2H), 3.47 (t, J=6.8Hz, 2H), 3.34 (s, 2H), 3.26-3.21 (m, 3H), 2.48 (s, 3H), 2.25-2.21 (m,1H), 2.12-2.06 (m, 1H), 1.82-1.77 (m, 2H), 1.60-1.56 (m, 2H), 1.45-1.36(m, 4H), 1.04 (s, 9H). MS (ESI); m/z: [M+H]⁺ calcd for C₄₀H₆₄IN₄O₁₀S:919.3, found 919.1.

N-(2-chloro-6-methylphenyl)-2-((6-(4-((S)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19,22-hexaoxa-4-azaoctacosan-28-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-2-2-2-2-2-2-VHL

Yield (68%); ¹H NMR (500 MHz, CD₃OD) δ 8.87 (s, 1H), 8.14 (s, 1H),7.47-7.39 (m, 4H), 7.34 (d, J=7.1 Hz, 1H), 7.26-7.20 (m, 2H), 6.00 (s,1H), 4.68 (bs, 1H), 4.59-4.50 (m, 3H), 4.37-4.33 (m, 1H), 4.08-4.00 (m,2H), 3.87 (d, J=11.0 Hz, 1H), 3.80 (dd, J=11.0 Hz, J=3.8 Hz, 1H),3.70-3.61 (m, 22H), 3.58-3.55 (m, 2H), 3.46 (t, J=6.8 Hz, 2H), 2.58-2.51(m, 4H), 2.50-2.45 (m, 6H), 2.42-2.39 (m, 2H), 2.32 (s, 3H), 2.25-2.20(m, 1H), 2.12-2.06 (m, 1H), 1.60-1.53 (m, 4H), 1.42-1.32 (m, 4H), 1.04(s, 9H). ¹³C NMR (151 MHz, CD₃OD) δ 173.0, 170.0, 169.8, 168.4, 166.8,165.9, 163.0, 161.8, 157.1, 152.1, 140.7, 138.9, 136.2, 135.8, 132.9,132.8, 131.6, 128.7, 128.1, 126.9, 125.4, 124.4, 118.1, 116.4, 82.5,71.2, 70.8, 70.4, 70.3, 70.2, 70.16, 70.14, 70.12, 69.8, 58.2, 52.4,49.2, 48.1, 43.3, 30.7, 29.2, 26.9, 25.9, 25.7, 24.2, 22.2, 17.29,17.28. MS (ESI); m/z: [M+H] calcd for C₆₀H₈₅ClN₁₁O₁₁S₂: 1234.5, found1234.8.

(2S,4R)-1-((S)-2-(tert-butyl)-27-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:IMA-6-2-2-2-2-2-2-VHL

Yield (54%); ¹H NMR (500 MHz, CDCl₃/d₆DMSO) δ 9.23 (d, J=2.0 Hz, 1H),8.69-8.66 (m, 2H), 8.57 (s, 1H), 8.51-8.48 (m, 2H), 8.20 (s, 1H), 7.84(d, J=8.0 Hz, 2H), 7.42-7.28 (m, 10H), 7.20-7.16 (m, 2H), 7.09 (s, 1H),4.70 (t, J=7.9 Hz, 1H), 4.56-4.48 (m, 3H), 4.33 (d, J=15.1 Hz, J=5.3 Hz,1H), 4.03-3.98 (m, 3H), 3.64-3.53 (m, 23H), 3.42 (t, J=6.5 Hz, 2H),2.72-2.55 (m, 7H), 2.52-2.45 (m, 6H), 2.33 (s, 3H), 2.14-2.08 (m, 1H),1.58-1.51 (m, 4H), 1.36-1.29 (m, 6H), 0.94 (s, 9H). ¹³C NMR (151 MHz,CDCl₃/d₆DMSO) δ 173.2, 170.8, 170.3, 165.5, 162.7, 160.6, 159.0, 151.4,150.3, 148.46, 148.41, 141.9, 138.1, 137.7, 136.6, 134.9, 134.0, 132.7,131.6, 130.9, 130.7, 129.4, 129.3, 128.1, 127.2, 124.5, 123.7, 115.5,113.4, 108.3, 71.2, 71.1, 70.5, 70.52, 70.51, 70.47, 70.45, 70.38, 70.3,69.9, 62.2, 58.5, 58.3, 57.0, 56.7, 54.7, 52.9, 52.0, 43.2, 36.0, 35.1,29.4, 27.1, 25.9, 17.7, 16.0. MS (ESI); m/z: [M+H]⁺ calcd forC₆₈H₉₂N₁₁O₁₁S: 1270.6, found 1270.2.

(2S,4R)-1-((S)-2-(6-((5-((6-chlorohexyl)oxy)pentyl)oxy)hexanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Yield (72%); ¹H NMR (500 MHz, CD₃OD) δ 8.87 (s, 1H), 7.45 (d, J=7.8 Hz,2H), 7.41 (d, J=7.8 Hz, 2H), 4.83 (s, 2H), 4.63 (bs, 1H), 4.58-4.49 (m,3H), 4.37-4.43 (m, 1H), 3.90 (d, J=11.0 Hz, 1H), 3.80 (dd, J=11.0 Hz,J=3.8 Hz, 1H), 3.54 (t, J=6.8 Hz, 2H), 3.43-3.38 (m, 6H), 2.47 (s, 3H),2.32-2.19 (m, 3H), 2.11-2.06 (m, 1H), 1.78-1.72 (m, 2H), 1.65-1.54 (m,10H), 1.47-1.35 (m, 8H), 1.03 (s, 9H). ¹³C NMR (151 MHz, CD₃OD) δ 174.5,173.1, 170.9, 151.4, 147.6, 138.9, 132.0, 130.0, 128.9, 127.0, 70.43,70.41, 70.31, 70.29, 69.7, 59.4, 59.3, 57.5, 56.6, 44.3, 42.4, 42.3,37.5, 35.2, 35.1, 32.3, 29.2, 29.1, 29.0, 26.3, 25.6, 25.5, 25.4, 22.5,17.3, 14.4. MS (ESI); m/z: [M+H]⁺ calcd for C₃₉H₆₂ClN₄O₆S: 749.4, found749.2.

(2S,4R)-4-hydroxy-1-((S)-2-(6-((5-((6-iodohexyl)oxy)pentyl)oxy)hexanamido)-3,3-dimethylbutanoyl)-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

Yield (87%); ¹H NMR (500 MHz, CD₃OD) δ 8.87 (s, 1H), 7.45 (d, J=7.8 Hz,2H), 7.41 (d, J=7.8 Hz, 2H), 4.84 (s, 2H), 4.65-4.62 (m, 1H), 4.58-4.49(m, 3H), 4.37-4.43 (m, 1H), 3.90 (d, J=11.0 Hz, 1H), 3.80 (dd, J=11.0Hz, J=3.8 Hz, 1H), 3.43-3.38 (m, 6H), 3.22 (t, J=7.0 Hz, 2H), 2.47 (s,3H), 2.32-2.19 (m, 3H), 2.11-2.06 (m, 1H), 1.82-1.77 (m, 2H), 1.65-1.54(m, 10H), 1.47-1.35 (m, 8H), 1.03 (s, 9H). MS (ESI); m/z: [M+H]⁺ calcdfor C₃₉H₆₂IN₄O₆S: 841.3, found 841.0

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-((5-((6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-5-6-VHL

Yield (54%); ¹H NMR (500 MHz, CD₃OD/CDCl₃) δ 8.55 (s, 1H), 7.91 (s, 1H),7.22-7.14 (m, 4H), 7.06-6.99 (m, 2H), 6.91 (d, J=9.3 Hz, 1H), 5.78 (s,1H), 4.43-4.34 (m, 4H), 4.20 (dd, J=15.0 Hz, J=4.9 Hz, 1H), 3.77 (d,J=11.0 Hz, 1H), 3.55 (dd, J=11.0 Hz, J=3.8 Hz, 1H), 3.47 (bs, 4H),3.28-3.21 (m, 8H), 2.39 (bs, 4H), 2.33 (bs, 6H), 2.25-2.22 (m, 2H), 2.16(s, 3H), 2.11-2.02 (m, 4H), 1.49-1.37 (m, 12H), 1.26-1.17 (m, 8H), 0.83(s, 9H). ¹³C NMR (151 MHz, CD₃OD/CDCl₃) δ 174.2, 174.1, 172.1, 171.1,166.3, 163.9, 163.0, 161.4, 156.9, 150.9, 147.8, 140.5, 138.7, 138.3,132.6, 132.5, 132.0, 130.3, 129.3, 128.9, 128.1, 127.7, 127.0, 125.5,83.0, 70.8, 70.7, 70.6, 69.7, 59.2, 59.6, 57.5, 57.4, 56.9, 52.6, 49.5,43.6, 42.8, 36.9, 36.0, 35.5, 29.4, 29.3, 29.2, 27.3, 26.3, 26.2, 25.9,25.7, 25.4, 25.2, 22.6, 18.4, 15.4. MS (ESI); m/z: [M+H]⁺ calcd forC₅₉H₈₃ClN₁₁O₇S₂: 1156.5, found 1156.2.

(2S,4R)-1-((S)-3,3-dimethyl-2-(6-((5-((6-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:IMA-6-5-6-VHL

Yield (47%); ¹H NMR (500 MHz, CDCl₃) δ 9.23 (d, J=2.0 Hz, 1H), 8.69 (d,J=3.4 Hz, 1H), 8.67 (s, 1H), 8.56 (s, 1H), 8.52-8.48 (m, 2H), 8.08 (s,1H), 7.84 (d, J=8.1 Hz, 2H), 7.45-7.40 (m, 3H), 7.35-7.29 (m, 6H),7.21-7.17 (m, 2H), 7.14 (s, 1H), 6.17 (d, J=8.6 Hz, 1H), 4.70 (t, J=7.8Hz, 1H), 4.58-4.49 (m, 3H), 4.31 (d, J=14.9 Hz, J=5.1 Hz, 1H), 4.03 (d,J=11.2 Hz, 1H), 3.59-3.55 (m, 5H), 3.39-3.34 (m, 8H), 2.60-2.44 (m,10H), 2.40-2.34 (m, 5H), 2.20-2.08 (m, 4H), 1.62-1.43 (m, 12H),1.40-1.29 (m, 8H), 0.91 (s, 9H). ¹³C NMR (151 MHz, CDCl₃) δ 173.5,171.9, 170.7, 165.5, 162.7, 160.6, 159.0, 151.4, 150.3, 148.46, 148.44,142.3, 138.0, 137.7, 136.6, 134.9, 133.9, 132.7, 131.6, 130.9, 130.8,129.5, 129.3, 128.1, 127.0, 124.6, 123.7, 115.5, 113.5, 108.3, 70.8,70.76, 70.71, 70.5, 69.9, 62.5, 58.6, 57.4, 56.7, 53.1, 43.2, 36.4,35.8, 34.9, 29.6, 29.54, 29.51, 29.3, 27.4, 26.4, 26.1, 25.8, 25.3,22.8, 17.7, 16.0. MS (ESI); m/z: [M+H]⁺ calcd for C₆₇H₉₀N₁₁O₇S: 1192.6,found 1192.2.

Synthesis of Bosutinib with VHL Ligand

tert-butyl-6-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)ethoxy)hexanoate

To a solution of4-(2,4-dichloro-5-methoxy-anilino)-6-methoxy-7-(3-piperazin-1-ylpropoxy)quinoline-3-carbonitrile(40 mg, 0.08 mmol) and K₂CO₃ (32.11 mg, 0.23 mmol) in DMF (2 ml) wasadded tert-butyl 6-[2-[2-(6-chlorohexoxy)ethoxy]ethoxy]hexanoate (36.71mg, 0.09 mmol) and the resulting solution stirred for 16 h at 100° C.Then the mixture was cooled down to rt. After filtration, the solventwas evaporated and the residue subjected to Prep TLC purification(MeOH/DCM:95/5) to give 11 mg (16.2%) of tert-butyl6-[2-[2-[6-[4-[3-[[3-cyano-4-(2,4-dichloro-5-methoxy-anilino)-6-methoxy-7-quinolyl]oxy]propyl]piperazin-1-yl]hexoxy]ethoxy]ethoxy]hexanoateas a yellow oil. ¹H NMR (500 MHz, CDCl₃/TMS) δ 8.69 (s, 1H), 7.48 (s,1H), 7.42 (s, 1H), 6.92 (s, 1H), 6.79 (s, 1H), 6.48 (s, 1H), 4.25 (t,J=6.4 Hz, 2H), 3.77 (s, 3H), 3.66 (s, 3H), 3.65-3.61 (m, 4H), 3.58-3.55(m, 4H), 346-3.41 (m, 4H), 2.62-2.42 (m, 12H), 2.19 (t, J=7.4 Hz, 2H),2.13-2.08 (m, 2H), 1.61-1.54 (m, 8H), 1.43 (s, 9H), 1.40-1.28 (m, 6H).¹³C NMR (151 MHz, CDCl₃/TMS) δ 173.1, 154.3, 153.9, 150.3, 149.8, 147.7,147.5, 136.9, 130.5, 118.4, 117.2, 116.4, 114.8, 109.9, 105.4, 101.1,94.2, 79.9, 71.3, 71.2, 70.61, 70.60, 70.1, 67.6, 58.4, 56.5, 56.1,54.7, 52.9, 35.5, 29.5, 29.3, 28.1, 27.3, 26.1, 25.9, 25.6, 24.9. MS(ESI); m/z: [M+H]⁺ calcd for C₄₅H₆₆Cl₂N₅O₈: 874.4, found 874.2.

6-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)ethoxy)hexanoicacid

A solution oftert-butyl-6-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)ethoxy)hexanoate(12 mg, 0.01 mmol) in DCM/TF (1 mL) was stirred at room temperature for2 h. The solvent was evaporated to give 11.23 mg (100%) of the desiredproduct which was carried to the next step without further purification.

(2S,4R)-1-((S)-2-(tert-butyl)-22-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:BOS-6-2-2-6-VHL

To a solution of6-[2-[2-[6-[4-[3-[[3-cyano-4-(2,4-dichloro-5-methoxy-anilino)-6-methoxy-7-quinolyl]oxy]propyl]piperazin-1-yl]hexoxy]ethoxy]ethoxy]hexanoic acid (11 mg, 0.01 mmol) in DMF (1 mL) was added HATU (10.22 mg,0.03 mmol) and the resulting solution stirred for 10 minutes at rt,after which((2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide;hydrochloride (6.27 mg, 0.01 mmol) andDIEA (0.01 ml, 0.07 mmol) were added respectively. The resulting mixturewas stirred at room temperature for 16 h at rt. The product wasextracted twice with Ac₂O then purified by Prep TLC DCM/(MeOH with 2%ammonia) 95:5 to give 7.9 mg (47.8%) of the desired product as a yellowoil. ¹H NMR (500 MHz, CDCl₃) δ 8.69 (s, 1H), 8.68 (s, 1H), 7.49 (s, 1H),7.43 (s, 1H), 7.39-7.32 (m, 4H), 7.29 (t, J=6.4 Hz, 1H), 6.95 (s, 1H),6.86 (s, 1H), 6.52 (s, 1H), 6.10 (d, J=8.3 Hz, 1H), 4.72 (t, J=7.8 Hz,1H), 4.60-4.49 (m, 3H), 4.34 (dd, J=14.0 Hz, J=4.9 Hz, 1H), 4.26 (t,J=6.4 Hz, 2H), 4.07 (d, J=11.2 Hz, 1H), 3.79 (s, 3H), 3.70-3.55 (m,22H), 3.45-3.42 (m, 4H), 2.75-2.62 (m, 8H), 2.57-2.53 (m, 2H), 2.51 (s,3H), 2.21-2.11 (m, 5H), 1.62-1.53 (m, 8H), 1.37-1.29 (m, 6H), 0.92 (s,9H). ¹³C NMR (151 MHz, CDCl₃) δ 173.6, 171.9, 170.6, 154.3, 153.8,150.3, 150.2, 149.8, 148.5, 147.6, 147.5, 138.0, 136.9, 132.9, 131.5,130.9, 130.5, 129.6, 129.5, 128.3, 128.1, 118.5, 117.8, 116.4, 114.8,109.8, 105.7, 101.1, 93.9, 71.2, 71.6, 70.6, 70.3, 70.0, 69.9, 67.4,63.9, 58.4, 58.3, 57.4, 56.7, 56.5, 56.1, 54.6, 52.7, 43.2, 36.4, 35.8,34.8, 29.5, 29.2, 27.1, 26.4, 25.9, 25.8, 25.7, 25.3, 16.0. MS (ESI);m/z: [M+H]⁺ calcd for C₆₃H₈₆Cl₂N₉O₁₀S: 1230.5, found 1230.1.

tert-butyl-2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetate

Yield (30%); ¹H NMR (500 MHz, CDCl₃) δ 8.70 (s, 1H), 7.49 (s, 1H), 7.42(s, 1H), 6.90 (s, 1H), 6.75 (s, 1H), 6.46 (s, 1H), 4.25 (t, J=6.8 Hz,2H), 4.02 (s, 2H), 3.77 (s, 3H), 3.71-3.67 (m, 2H), 3.66 (s, 3H),3.62-3.58 (m, 2H), 3.45 (t, J=6.8 Hz, 2H), 2.65-2.52 (m, 12H), 2.15-2.08(m, 2H), 1.61-1.55 (m, 2H), 1.50-1.32 (m, 15H). ¹³C NMR (151 MHz, CDCl₃)δ 169.7, 154.2, 153.9, 150.3, 149.7, 147.7, 147.4, 136.9, 130.5, 118.2,117.0, 116.4, 114.8, 109.9, 105.2, 101.0, 99.3, 81.5, 71.4, 70.7, 70.0,69.0, 67.7, 59.4, 56.5, 56.1, 54.8, 53.1, 53.0, 52.9, 52.8, 29.5, 28.1,28.0, 27.4, 26.2, 26.0. MS (ESI); m/z: [M+H]⁺ calcd for C₃₉H₅₄C₂N₅O₇:774.3, found 774.2.

(2S,4R)-1-((S)-2-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:BOS-6-2-2-VHL

Yield (55%); ¹H NMR (500 MHz, CD₃OD/CDCl₃) δ 8.67 (bs, 2H), 7.48 (s,1H), 7.44 (s, 1H), 7.38-7.29 (m, 6H), 6.95 (s, 1H), 6.86 (s, 1H), 6.53(s, 1H), 4.71 (t, J=7.8 Hz, 1H), 4.56-4.47 (m, 3H), 4.35 (dd, J=14.0 Hz,J=4.1 Hz, 1H), 4.26 (bs, 2H), 4.08-3.94 (m, 3H), 3.79 (s, 3H), 3.74-3.55(m, 10H), 3.50-3.41 (m, 4H), 3.18-3.12 (m, 2H), 2.77-2.47 (m, 11H),2.16-2.02 (m, 3H), 1.62-1.51 (m, 4H), 1.48-1.31 (m, 4H), 0.95 (s, 9H).¹³C NMR (151 MHz, CD₃OD/CDCl₃) δ 171.3, 170.8, 170.5, 154.3, 153.7,150.3, 150.2, 149.9, 148.4, 147.6, 147.5, 138.0, 136.8, 131.6, 130.9,130.5, 129.5, 128.1, 120.6, 118.5, 117.5, 116.5, 114.7, 109.8, 105.9,101.1, 71.3, 71.1, 70.3, 70.1, 69.9, 67.4, 58.5, 58.3, 57.2, 56.6, 56.5,56.1, 55.5, 54.6, 52.9, 50.8, 43.5, 43.2, 35.9, 34.8, 29.2, 26.4, 25.9,18.0, 16.1, 12.6. MS (ESI); m/z: [M+H]⁺ calcd for C₅₇H₇₄C₂N₉O₉S: 1130.4,found 1130.1.

tert-butyl-24-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxatetracosanoate

Yield (41%); ¹H NMR (500 MHz, CDCl₃) δ 8.67 (s, 1H), 7.47 (s, 1H), 7.41(s, 1H), 6.95 (s, 1H), 6.88 (s, 1H), 6.51 (s, 1H), 4.24 (t, J=6.8 Hz,2H), 3.99 (s, 2H), 3.78 (s, 3H), 3.70-3.60 (m, 21H), 3.57-3.54 (m, 2H),3.42 (t, J=6.8 Hz, 2H), 2.58-2.45 (m, 10H), 2.33-2.30 (m, 2H), 2.12-2.07(m, 2H), 1.58-1.53 (m, 2H), 1.49-1.43 (m, 11H), 1.35-1.28 (m, 4H). ¹³CNMR (151 MHz, CDCl₃) δ 169.6, 154.2, 153.9, 150.3, 149.8, 147.6, 136.9,130.5, 118.5, 117.5, 116.5, 114.7, 109.8, 105.8, 101.1, 93.8, 81.5,71.4, 70.7, 70.6, 70.54, 70.52, 70.51, 70.0, 68.9, 67.7, 58.7, 56.5,56.1, 54.8, 53.2, 53.1, 29.5, 28.1, 27.4, 26.7, 26.2, 26.0. MS (ESI);m/z: [M+H]⁺ calcd for C₄₇H₇₀C₂N₅O₁₁: 950.4, found 950.2.

(2S,4R)-1-((S)-2-(tert-butyl)-27-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:BOS-6-2-2-2-2-2-2-VHL

Yield (74%); ¹H NMR (500 MHz, CD₃OD/CDCl₃) δ 8.66 (s, 1H), 8.62 (s, 1H),7.47-7.42 (m, 2H), 7.38 (s, 1H), 7.35-7.30 (m, 4H), 7.01 (s, 1H), 6.57(s, 1H), 4.68 (t, J=7.8 Hz, 1H), 4.54-4.49 (m, 3H), 4.35 (dd, J=14.0 Hz,J=4.9 Hz, 1H), 4.23 (t, J=5.8 Hz, 2H), 4.03-3.94 (m, 3H), 3.74 (s, 3H),3.69 (s, 3H), 368-350 (m, 20H), 3.41 (t, J=6.8 Hz, 2H), 3.11-3.07 (m,1H), 2.62-2.37 (m, 17H), 2.16-2.08 (m, 3H), 1.57-1.49 (m, 4H), 1.40-1.33(m, 4H), 0.94 (s, 9H). ¹³C NMR (151 MHz, CD₃OD/CDCl₃) δ 171.2, 170.9,170.5, 154.3, 153.7, 150.4, 150.2, 149.9, 148.3, 147.9, 147.2, 138.2,136.7, 131.7, 130.7, 130.5, 129.4, 128.0, 118.9, 118.3, 116.5, 114.5,109.6, 106.7, 101.1, 92.9, 71.3, 70.9, 70.43, 70.40, 70.36, 70.33,70.30, 70.2, 69.9, 69.8, 67.5, 58.7, 58.4, 57.0, 56.8, 56.6, 56.1, 54.9,54.7, 52.8, 52.5, 50.6, 43.1, 43.0, 36.3, 35.3, 29.3, 26.3, 25.8, 17.9,15.9, 12.7. MS (ESI); m/z: [M+H]⁺ calcd for C₆₅H₉₀C₂N₉O₁₃S: 1306.6,found 1306.4.

tert-butyl-6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanoate

Yield (68%); ¹H NMR (500 MHz, CDCl₃) δ 8.66 (s, 1H), 7.48 (s, 1H), 7.41(s, 1H), 6.97 (bs, 2H), 6.52 (s, 1H), 4.24 (t, J=5.8 Hz, 2H), 3.78 (s,3H), 3.68 (s, 3H), 3.40-3.35 (m, 8H), 2.82-2.66 (m, 12H), 2.20 (t, J=7.8Hz, 2H), 2.11 (bs, 2H), 1.71-1.50 (m, 12H), 1.43 (s, 9H), 1.40-1.30 (m,8H). ¹³C NMR (151 MHz, CDCl₃) δ 173.2, 154.3, 153.6, 150.2, 149.9,147.7, 147.4, 136.8, 130.5, 118.7, 117.7, 116.4, 114.7, 109.8, 106.1,101.1, 93.7, 79.9, 70.8, 70.7, 70.6, 70.5, 68.9, 57.4, 56.5, 56.1, 54.1,51.9, 50.6, 35.5, 29.6, 29.5, 29.48, 29.4, 28.1, 26.7, 25.8, 25.6, 24.9,22.8. MS (ESI); m/z: [M+H]⁺ calcd for C₄₆H₆₈Cl₂N₅O₇: 872.4, found 872.2.

(2S,4R)-1-((S)-2-(6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide:BOS-6-5-6-VHL

Yield (38%); ¹H NMR (500 MHz, CDCl₃/TMS) δ 8.69 (s, 1H), 8.68 (s, 1H),7.49 (s, 1H), 7.42 (s, 1H), 7.39-7.32 (m, 4H), 7.29 (t, J=6.4 Hz, 1H),6.95 (s, 1H), 6.87 (s, 1H), 6.51 (s, 1H), 6.09 (d, J=8.8 Hz, 1H), 4.72(t, J=7.8 Hz, 1H), 4.59-4.49 (m, 3H), 4.33 (dd, J=14.0 Hz, J=4.9 Hz,1H), 4.26 (t, J=7.4 Hz, 2H), 4.07 (d, J=11.2 Hz, 1H), 3.79 (s, 3H),3.74-3.59 (m, 12H), 3.40-3.36 (m, 6H), 2.75-2.67 (m, 8H), 2.57-2.53 (m,2H), 2.51 (s, 3H), 2.21-2.18 (m, 2H), 2.16-2.08 (m, 3H), 1.62-1.52 (m,12H), 1.41-1.32 (m, 8H), 0.92 (s, 9H). ¹³C NMR (151 MHz, CDCl₃/TMS) δ171.6, 171.9, 170.6, 154.3, 153.7, 150.3, 150.2, 149.8, 148.5, 147.6,147.5, 138.0, 136.9, 131.6, 130.9, 130.5, 129.5, 128.1, 118.6, 117.5,116.4, 114.8, 109.8, 105.8, 101.1, 93.9, 70.78, 70.76, 70.7, 70.5, 69.9,67.4, 58.4, 58.2, 57.4, 56.6, 56.5, 56.1, 54.5, 52.7, 43.2, 36.4, 35.8,34.8, 29.7, 29.6, 29.53, 29.51, 29.3, 27.1, 26.4, 25.9, 25.8, 25.3,22.8, 16.0. MS (ESI); m/z: [M+H]⁺ calcd for C₆₄H₈₈Cl₂N₉O₉S: 1228.6,found 1229.0.

Synthesis of Dasatinib/Bosutinib/Imatinib with Pomalidomide

6-(2-(2-((6-chlorohexyl)oxy)ethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide

(6-[2-[2-(6-chlorohexoxy)ethoxy]ethoxy]hexanoyl chloride (110 mg, 0.31mmol) was dissolved in THE (2 ml). To this solution was added4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione (84.12 mg, 0.31mmol). The resulting suspension was heated to reflux for 4 hours. Thesolvent was evaporated in vacuo and the resulting solid was purified byflash chromatography (50/50 to 0/100 hexane/ethyl acetate) to give alight yellow solid 157 mg (85.8%) of the desired product. ¹H NMR (500MHz, Chloroform-d) δ 9.41 (s, 1H), 8.82 (d, J=7.8 Hz, 1H), 8.45 (s, 1H),7.70 (t, J=7.8 Hz, 1H), 7.54 (d, J=7.8 Hz, 1H), 4.96 (dd, J=12.0 Hz,J=6.5 Hz, 1H), 3.66-3.61 (m, 4H), 3.60-3.57 (m, 4H), 3.52 (t, J=6.3 Hz,2H), 3.50-3.43 (m, 4H), 2.91 (d, J=13.6 Hz, 1H), 2.85-2.72 (m, 2H), 2.46(t, J=7.3 Hz, 2H), 2.21-2.12 (m, 1H), 1.80-1.73 (m, 4H), 1.67-1.55 (m,4H), 1.50-1.34 (m, 6H). ¹³C NMR (151 MHz, Chloroform-d) δ 172.2, 170.9,169.1, 167.9, 166.7, 137.8, 136.4, 131.1, 125.2, 118.4, 115.2, 71.2,71.0, 70.6, 70.0, 49.2, 45.0, 37.9, 32.5, 31.4, 29.4, 29.3, 26.7, 25.7,25.4, 25.0, 22.6. MS (ESI); m/z: [M+H]⁺ calcd for C₂₉H₄₁ClN₃O: 594.2,found 594.1.

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-6-(2-(2-((6-iodohexyl)oxy)ethoxy)ethoxy)hexanamide

To a solution of6-[2-[2-(6-chlorohexoxy)ethoxy]ethoxy]-N-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]hexanamide(130 mg, 0.22 mmol) in Acetone (10 ml) was added NaI (164 mg, 1.09mmol). The reaction mixture was stirred at reflux temperature for 24 h,then the solvent was removed under vacuum and crude product wasdissolved in EtOAc (15 mL) and an aqueous solution of Na₂SO₃ (10%, 10mL), the organic layer was separated, washed with water (10 mL), dried(Na₂SO₄) and evaporated under vacuum. The crude product was pure by NMR(>98% purity), 127 mg (83.8%) of the desired product. It was used in thenext step without any further purification. ¹H NMR (400 MHz,Chloroform-d) δ 9.41 (s, 1H), 8.82 (d, J=7.8 Hz, 1H), 8.10 (s, 1H), 7.70(t, J=7.8 Hz, 1H), 7.55 (d, J=7.8 Hz, 1H), 4.95 (dd, J=12.0 Hz, J=6.5Hz, 1H), 3.66-3.61 (m, 4H), 3.60-3.56 (m, 4H), 3.50-3.42 (m, 4H), 3.17(t, J=7.0 Hz, 2H), 2.95-2.70 (m, 3H), 2.46 (t, J=7.3 Hz, 2H), 2.21-2.12(m, 1H), 1.86-1.73 (m, 4H), 1.67-1.55 (m, 4H), 1.50-1.34 (m, 6H). MS(ESI); m/z: [M+H]⁺ calcd for C₂₉H₄₁IN₃O₈: 686.2, found 686.0.

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)ethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-2-2-6-CRBN

To a solution ofN-(2-chloro-6-methyl-phenyl)-2-[(2-methyl-6-piperazin-1-yl-pyrimidin-4-yl)amino]thiazole-5-carboxamide;hydrochloride(21.02 mg, 0.04 mmol) and DIEA (99.71 μl, 0.57 mmol) in DMF (1 ml) wasaddedN-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-4-yl]-6-[2-[2-(6-iodohexoxy)ethoxy]ethoxy]hexanamide(15 mg, 0.02 mmol) and the resulting solution stirred for 16 h at 80° C.Then the mixture was cooled down to rt. The solvent was evaporated andthe residue subjected to Prep TLC purification (2% ammonia in MeOH/DCM:10/90) to give 7.4 mg (33.8%) of the desired product as a foamy yellowsolid. ¹H NMR (500 MHz, CD₃OD) δ 8.63 (d, J=8.3 Hz, 1H), 8.13 (s, 1H),7.74 (t, J=8.3 Hz, 1H), 7.53 (d, J=7.3 Hz, 1H), 7.35 (d, J=7.3 Hz, 1H),7.27-7.20 (m, 2H), 5.97 (s, 1H), 5.11 (dd, J=12.0 Hz, J=6.5 Hz, 1H),3.66-3.53 (m, 10H), 3.51-3.45 (m, 4H), 2.90-2.83 (m, 4H), 2.78-2.68 (m,2H), 2.58 (bs, 2H), 2.50 (t, J=7.3 Hz, 2H), 2.45 (s, 3H), 2.42 (m, 2H),2.32 (s, 3H), 2.19-2.12 (m, 1H), 1.76 (quint, J=7.3 Hz, 2H), 1.66-1.55(m, 6H), 1.50-1.34 (m, 7H). ¹³C NMR (151 MHz, CD₃OD) δ 173.0, 172.9,169.9, 168.5, 166.8, 166.0, 163.7, 163.0, 161.8, 157.1, 140.7, 138.9,136.8, 135.7, 132.9, 132.8, 131.5, 128.7, 128.1, 126.9, 125.9, 125.4,125.2, 117.9, 116.5, 82.5, 70.7, 71.6, 70.2, 69.8, 58.2, 52.4, 49.1,48.1, 43.3, 36.9, 30.7, 29.2, 28.9, 26.9, 25.9, 25.7, 25.4, 24.4, 24.2,22.2, 17.3. MS (ESI); m/z: [M+H]⁺ calcd for C₄₉H₆₂ClN₁₀O₉S: 1001.4,found 1001.1.

6-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide:BOS-6-2-2-6-CRBN

Yield (55%); ¹H NMR (500 MHz, CDCl₃) δ 9.41 (s, 1H), 8.81 (d, J=8.8 Hz,1H), 8.71 (s, 1H), 7.70 (t, J=7.8 Hz, 1H), 7.73 (d, J=7.3 Hz, 1H), 7.48(s, 1H), 7.45 (s, 1H), 6.93 (s, 1H), 6.82 (s, 1H), 6.49 (s, 1H), 4.92(dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.25 (t, J=6.8 Hz, 2H), 3.78 (s, 3H),3.67 (s, 3H), 3.65-3.61 (m, 4H), 3.59-3.55 (m, 4H), 3.50-3.42 (m, 4H),2.96-2.40 (m, 16H), 2.21-2.08 (m, 3H), 1.77 (quint, J=7.3 Hz, 2H),1.66-1.42 (m, 9H), 1.40-1.27 (m, 4H). ¹³C NMR (151 MHz, CDCl₃) δ 172.2,171.1, 169.2, 168.2, 166.7, 154.3, 153.9, 150.3, 149.8, 147.6, 147.5,137.8, 136.9, 136.4, 131.1, 130.6, 125.2, 118.4, 118.3, 117.2, 116.4,115.2, 114.8, 109.8, 105.5, 101.1, 99.1, 71.3, 70.9, 70.6, 70.1, 70.0,67.5, 58.4, 56.5, 56.1, 54.6, 52.8, 49.3, 37.9, 31.5, 29.5, 29.3, 27.3,26.0, 25.9, 25.7, 25.0, 22.7. MS (ESI); m/z: [M+H] calcd forC₅₄H₆₇C₂N₈O₁₁: 1073.4, found 1073.1.

4-((4-(6-(2-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)ethoxy)ethoxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide:IMA-6-2-2-6-CRBN

Yield (63%); ¹H NMR (500 MHz, CDCl₃) δ 9.40 (s, 1H), 9.24 (s, 1H), 8.80(d, J=8.1 Hz, 1H), 6.69 (dd, J=4.6 Hz, J=1.5 Hz, 1H), 8.53 (s, 1H),8.51-8.48 (m, 2H), 8.07 (s, 1H), 7.83 (d, J=8.1 Hz, 2H), 7.68 (t, J=8.3Hz, 1H), 7.51 (d, J=7.3 Hz, 1H), 7.42-7.39 (m, 3H), 7.32 (dd, J=7.3 Hz,J=1.5 Hz, 1H), 7.20-7.15 (m, 3H), 4.92 (dd, J=12.5 Hz, J=5.4 Hz, 1H),3.63-3.62 (m, 4H), 3.58-3.55 (m, 6H), 3.47-3.42 (m, 6H), 2.89-2.68 (m,4H), 2.59 (bs, 4H), 2.44 (t, J=7.8 Hz, 2H), 2.41-2.37 (m, 2H), 2.33 (s,3H), 2.16-2.12 (m, 1H), 1.76 (quint, J=7.3 Hz, 2H), 1.65-1.14 (m, 9H),1.37-1.25 (m, 4H). ¹³C NMR (151 MHz, CDCl₃) δ 172.1, 171.3, 169.1,168.3, 166.8, 165.4, 162.7, 160.5, 158.9, 151.4, 148.4, 142.0, 137.8,137.7, 136.6, 136.4, 135.0, 133.9, 132.6, 131.1, 130.7, 129.4, 127.1,125.2, 124.5, 123.7, 118.4, 115.5, 115.2, 113.4, 108.3, 71.3, 71.0,70.6, 70.1, 70.0, 62.4, 58.4, 52.9, 52.5, 50.8, 49.3, 37.9, 31.5, 29.5,29.3, 27.3, 26.2, 25.9, 25.7, 25.0, 22.7, 17.7. MS (ESI); m/z: [M+H]⁺calcd for C₅₇H₆₉N₁₀O₉: 1037.5, found 1037.4.

2-(2-((6-chlorohexyl)oxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide

Yield (63%); ¹H NMR (500 MHz, Chloroform-d) δ 10.46 (s, 1H), 8.87 (d,J=8.3 Hz, 1H), 8.23 (s, 1H), 7.72 (t, J=8.3 Hz, 1H), 7.58 (d, J=8.3 Hz,1H), 4.96 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.22 (s, 2H), 3.84-3.81 (m,2H), 3.74-3.70 (m, 2H), 3.54-3.46 (m, 4H), 2.95-2.71 (m, 3H), 2.19-2.13(m, 1H), 1.74 (quint, J=7.3 Hz, 2H), 1.58 (quint, J=7.3 Hz, 2H),1.46-1.32 (m, 4H). ¹³C NMR (151 MHz, Chloroform-d) δ 170.8, 169.6,168.3, 167.8, 166.7, 136.8, 136.3, 131.1, 125.3, 118.9, 116.1, 71.6,71.4, 71.1, 70.2, 49.2, 45.0, 32.4, 31.9, 29.4, 26.6, 25.4, 22.6. MS(ESI); m/z: [M+H]⁺ calcd for C₂₃H₂₉ClN₃O₇: 494.2, found 494.0.

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-2-(2-((6-iodohexyl)oxy)ethoxy)acetamide

Yield (85%); ¹H NMR (500 MHz, Chloroform-d) δ 10.45 (s, 1H), 8.86 (d,J=7.8 Hz, 1H), 8.36 (s, 1H), 7.72 (t, J=7.8 Hz, 1H), 7.57 (d, J=7.8 Hz,1H), 4.95 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.22 (s, 2H), 3.84-3.82 (m,2H), 3.73-3.70 (m, 2H), 3.47 (t, J=6.3 Hz, 2H), 3.15 (t, J=6.8 Hz, 2H),2.94-2.70 (m, 3H), 2.20-2.11 (m, 1H), 1.78 (quint, J=6.8 Hz, 2H), 1.57(quint, J=6.8 Hz, 2H), 1.43-1.30 (m, 4H). MS (ESI); m/z: [M+H]⁺ calcdfor C₂₃H₂₉N₃O₇: 586.1, found 586.0.

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-2-2-CRBN

Yield (57%); ¹H NMR (500 MHz, CD₃OD) δ 8.88 (d, J=8.3 Hz, 1H), 8.14 (s,1H), 7.78 (t, J=8.3 Hz, 1H), 7.57 (d, J=7.3 Hz, 1H), 7.35 (d, J=7.3 Hz,1H), 7.27-7.20 (m, 2H), 5.98 (s, 1H), 5.12 (dd, J=12.0 Hz, J=6.5 Hz,1H), 4.21 (s, 2H), 3.84-3.80 (m, 2H), 3.75-3.71 (m, 2H), 3.64-3.59 (m,4H), 3.50 (t, J=6.4 Hz, 2H), 2.92-2.86 (m, 1H), 2.79-2.70 (m, 2H),2.52-2.48 (m, 4H), 2.46 (s, 3H), 2.36-2.31 (m, 5H), 2.20-2.13 (m, 1H),1.58-1.46 (m, 4H), 1.40-1.26 (m, 4H). ¹³C NMR (151 MHz, CD₃OD) δ 173.1,170.2, 169.8, 168.4, 166.8, 166.0, 163.8, 163.0, 161.9, 157.1, 140.7,138.9, 136.2, 135.8, 132.9, 132.8, 131.6, 128.7, 128.1, 126.9, 125.4,124.5, 118.1, 116.5, 82.4, 71.3, 70.9, 70.6, 69.9, 58.1, 52.4, 49.2,48.4, 43.3, 30.8, 29.2, 26.9, 25.9, 25.7, 24.2, 22.2, 17.3. MS (ESI);m/z: [M+H]⁺ calcd for C₄₃H₅₀ClN₁₀O₈S: 901.3, found 901.1.

2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide:BOS-6-2-2-CRBN

Yield (62%); ¹H NMR (500 MHz, CDCl₃) δ 9.41 (s, 1H), 8.84 (d, J=8.3 Hz,1H), 8.71 (s, 1H), 7.71 (t, J=7.8 Hz, 1H), 7.57 (d, J=7.3 Hz, 1H), 7.48(s, 1H), 7.44 (s, 1H), 6.90 (s, 1H), 6.76 (s, 1H), 6.46 (s, 1H), 4.92(dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.29-4.13 (m, 4H), 3.81-3.71 (m, 7H),3.66 (s, 3H), 3.53 (t, J=6.4 Hz, 2H), 2.96-2.84 (m, 2H), 2.76-2.44 (m,13H), 2.17-2.04 (m, 3H), 1.64-1.42 (m, 4H), 1.40-1.26 (m, 4H). ¹³C NMR(151 MHz, CDCl₃) δ 171.8, 169.3, 168.6, 168.4, 167.0, 154.3, 153.9,150.3, 149.7, 147.7, 147.5, 137.0, 136.7, 136.1, 131.9, 130.6, 125.3,118.7, 118.3, 117.0, 116.4, 116.2, 114.8, 109.9, 105.4, 101.1, 94.4,71.7, 71.5, 70.9, 69.8, 67.6, 67.3, 58.0, 56.5, 56.1, 54.6, 52.7, 49.6,45.6, 39.4, 31.9, 29.2, 27.1, 26.1, 25.9, 22.5. MS (ESI); m/z: [M+H]⁺calcd for C₄₈H₅₅Cl₂NO₁₀: 973.3, found 1073.5.

4-((4-(6-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide:IMA-6-2-2-CRBN

Yield (50%); ¹H NMR (500 MHz, CDCl₃) δ 10.41 (s, 1H), 9.23 (s, 1H), 8.85(d, J=8.6 Hz, 1H), 8.70 (dd, J=4.6 Hz, J=1.5 Hz, 1H), 8.57 (s, 1H),8.53-8.50 (m, 2H), 8.04 (s, 1H), 7.83 (d, J=8.1 Hz, 2H), 7.68 (t, J=8.3Hz, 1H), 7.53 (d, J=7.3 Hz, 1H), 7.43-7.39 (m, 3H), 7.32 (d, J=8.1 Hz,1H), 7.21-7.15 (m, 2H), 7.06 (s, 1H), 4.88 (dd, J=13.0 Hz, J=5.1 Hz,1H), 4.21-4.12 (m, 2H), 3.81-3.66 (m, 4H), 3.57 (s, 2H), 3.53-3.48 (m,2H), 2.92-2.81 (m, 2H), 2.72-2.65 (m, 2H), 2.49 (bs, 4H), 2.41-2.30 (m,7H), 2.13-2.09 (m, 1H), 1.71-1.42 (m, 5H), 1.37-1.25 (m, 4H). ¹³C NMR(151 MHz, CDCl₃) δ 171.9, 169.3, 168.6, 168.4, 167.1, 165.4, 162.7,160.5, 159.0, 151.5, 148.4, 141.9, 137.7, 136.7, 136.6, 136.1, 134.9,133.9, 132.6, 131.4, 130.7, 129.5, 126.9, 125.4, 124.3, 123.7, 118.7,116.2, 115.3, 113.2, 108.3, 71.7, 71.6, 70.9, 69.5, 62.3, 58.0, 52.7,52.3, 49.6, 31.8, 29.2, 27.2, 25.8, 22.5, 17.7. MS (ESI); m/z: [M+H]⁺calcd for C₅₁H₅₇N₁₀O: 937.4, found 937.1.

24-chloro-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-3,6,9,12,15,18-hexaoxatetracosanamide

Yield (91%); ¹H NMR (500 MHz, Chloroform-d) δ 10.47 (s, 1H), 8.85 (s,1H), 8.83 (d, J=8.8 Hz, 1H), 7.70 (t, J=8.3 Hz, 1H), 7.53 (d, J=7.3 Hz,1H), 4.92 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.17 (s, 2H), 3.72-3.62 (m,16H), 3.56-3.49 (m, 4H), 3.45-3.40 (m, 4H), 2.90-2.69 (m, 3H), 2.16-2.10(m, 1H), 1.78-1.71 (m, 2H), 1.60-1.53 (m, 2H), 1.45-1.32 (m, 4H). ¹³CNMR (151 MHz, Chloroform-d) δ 171.0, 169.3, 168.4, 167.9, 166.8, 136.7,136.2, 131.3, 125.1, 118.7, 116.1, 71.6, 71.2, 71.1, 70.9, 70.8, 70.64,70.60, 70.55, 70.51, 70.49, 70.44, 70.42, 70.1, 70.0, 68.6, 49.2, 45.0,32.5, 31.4, 29.4, 26.6, 25.4, 22.6. MS (ESI); m/z: [M+H]⁺ calcd forC₃₁H₄₅ClN₃O₁₁: 670.3, found 670.1.

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-24-iodo-3,6,9,12,15,18-hexaoxatetracosanamide

Yield (80%); ¹H NMR (500 MHz, Chloroform-d) δ 10.48 (s, 1H), 8.84 (d,J=8.3 Hz, 1H), 8.76 (s, 1H), 7.71 (t, J=7.8 Hz, 1H), 7.56 (d, J=7.3 Hz,1H), 4.92 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.18 (s, 2H), 3.72-3.56 (m,20H), 3.48-3.41 (m, 2H), 3.16 (t, J=6.8 Hz, 2H), 2.90-2.69 (m, 3H),2.15-2.11 (m, 1H), 1.80-1.72 (m, 2H), 1.60-1.53 (m, 2H), 1.43-1.34 (m,4H). MS (ESI); m/z: [M+H]⁺ calcd for C₃₁H₄₅IN₃O₁₁: 762.2, found 762.0.

N-(2-chloro-6-methylphenyl)-2-((6-(4-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-1-oxo-3,6,9,12,15,18-hexaoxatetracosan-24-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-2-2-2-2-2-2-CRBN

Yield (25%); ¹H NMR (500 MHz, CD₃OD) δ 8.78 (d, J=8.3 Hz, 1H), 8.13 (s,1H), 7.77 (t, J=8.3 Hz, 1H), 7.57 (d, J=7.3 Hz, 1H), 7.35 (d, J=7.3 Hz,1H), 7.27-7.20 (m, 2H), 5.98 (s, 1H), 5.13 (dd, J=12.0 Hz, J=6.5 Hz,1H), 4.20 (s, 2H), 3.83-3.77 (m, 4H), 3.68-3.54 (m, 20H), 3.46 (t, J=6.4Hz, 2H), 2.92-2.85 (m, 1H), 279-2.70 (m, 2H), 2.61-2.54 (m, 4H),2.49-2.42 (m, 5H), 2.32 (s, 3H), 2.20-2.13 (m, 1H), 1.61-1.53 (m, 4H),1.43-1.32 (m, 4H). ¹³C NMR (151 MHz, CD₃OD) δ 173.1, 170.0, 169.8,168.4, 166.8, 166.0, 163.8, 161.8, 157.1, 140.7, 138.9, 136.2, 135.8,132.9, 132.8, 131.6, 128.7, 128.1, 126.9, 125.4, 124.4, 118.1, 116.4,82.5, 71.2, 70.8, 70.4, 70.3, 70.2, 70.16, 70.14, 70.12, 69.8, 58.2,52.4, 49.2, 48.1, 43.3, 30.7, 29.2, 26.9, 25.9, 25.7, 24.2, 22.2, 17.3.MS (ESI); m/z: [M+H]⁺ calcd for C₅₁H₆₆ClN₁₀O₁₂S: 1077.4, found 1077.8.

24-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-3,6,9,12,15,18-hexaoxatetracosanamide:BOS-6-2-2-2-2-2-2-CRBN

Yield (75%); ¹H NMR (500 MHz, CDCl₃) δ 9.48 (s, 1H), 8.83 (d, J=8.3 Hz,1H), 8.70 (s, 1H), 7.71 (t, J=8.3 Hz, 1H), 7.55 (d, J=7.3 Hz, 1H), 7.48(s, 1H), 7.44 (s, 1H), 6.92 (s, 1H), 6.81 (s, 1H), 6.49 (s, 1H), 4.93(dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.25 (t, J=6.8 Hz, 2H), 4.19 (s, 2H),3.80 (s, 3H), 3.78 (s, 3H), 3.71-3.51 (m, 20H), 3.43 (t, J=6.8 Hz, 2H),2.96-2.70 (m, 4H), 2.62-2.35 (m, 12H), 2.17-2.06 (m, 3H), 1.58-1.54 (m,4H), 1.35-1.28 (m, 4H). ¹³C NMR (151 MHz, CDCl₃) δ 171.1, 169.3, 168.4,168.1, 167.8, 154.3, 153.9, 150.3, 149.8, 147.6, 147.5, 136.9, 136.7,136.2, 131.4, 130.5, 125.2, 118.7, 118.4, 117.2, 116.4, 116.1, 114.8,109.8, 105.5, 101.1, 94.1, 71.6, 71.3, 70.9, 70.7, 70.6, 70.56, 70.54,70.52, 70.48, 70.47, 67.6, 56.5, 56.1, 54.7, 52.9, 50.8, 49.3, 31.5,29.5, 27.4, 26.1, 25.9, 22.7. MS (ESI); m/z: [M+H]⁺ calcd forC₅₆H₇₁C₂N₈O₁₄: 1149.4, found 1149.7.

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-24-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxatetracosanamide:IMA-6-2-2-2-2-2-2-CRBN

Yield (43%); ¹H NMR (500 MHz, CDCl₃) δ 10.47 (s, 1H), 9.24 (s, 1H), 8.83(d, J=8.6 Hz, 1H), 8.70 (d, J=4.4 Hz, 1H), 8.55 (s, 1H), 8.51 (d, J=4.4Hz, 2H), 7.98 (s, 1H), 7.83 (d, J=8.1 Hz, 2H), 7.70 (t, J=7.8 Hz, 1H),7.54 (d, J=7.1 Hz, 1H), 7.45-7.39 (m, 3H), 7.32 (d, J=7.6 Hz, 1H),7.21-7.16 (m, 2H), 7.10 (s, 1H), 4.92 (dd, J=12.0 Hz, J=5.1 Hz, 1H),4.21-4.16 (m, 2H), 3.79 (s, 4H), 3.74-3.55 (m, 18H), 3.42 (t, J=6.4 Hz,2H), 2.95-2.68 (m, 4H), 2.51 (bs, 8H), 2.34 (s, 3H), 2.13-2.10 (m, 1H),1.58-1.42 (m, 5H), 1.40-1.25 (m, 4H). ¹³C NMR (151 MHz, CDCl₃) δ 171.2,169.3, 168.4, 168.2, 166.8, 165.4, 162.7, 160.5, 159.0, 151.4, 148.5,142.3, 137.7, 136.7, 136.6, 136.2, 134.9, 133.9, 132.7, 131.3, 130.7,129.4, 127.0, 125.2, 124.3, 123.7, 118.7, 116.1, 115.3, 113.2, 108.3,71.6, 71.3, 70.9, 70.6, 70.57, 70.53, 70.48, 70.46, 70.0, 62.5, 58.5,53.0, 52.8, 49.3, 31.5, 29.5, 27.4, 25.9, 22.7, 17.7. MS (ESI); m/z:[M+H]⁺ calcd for C₅₉H₇₃N₁₀O₁₂: 1113.5, found 1113.4.

6-((5-((6-chlorohexyl)oxy)pentyl)oxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide

Yield (58%); ¹H NMR (500 MHz, Chloroform-d) δ 9.40 (s, 1H), 8.82 (d,J=8.5 Hz, 1H), 8.31 (s, 1H), 7.70 (t, J=7.8 Hz, 1H), 7.54 (d, J=7.3 Hz,1H), 4.95 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 3.52 (t, J=6.4 Hz, 2H),3.44-3.37 (m, 8H), 2.93-2.72 (m, 3H), 2.46 (t, J=7.6 Hz, 2H), 2.20-2.12(m, 1H), 1.88-1.73 (m, 4H), 1.67-1.53 (m, 8H), 1.50-1.34 (m, 8H). ¹³CNMR (151 MHz, Chloroform-d) δ 172.2, 170.7, 169.1, 167.8, 166.7, 137.9,136.4, 131.1, 125.3, 118.4, 115.2, 70.8, 70.7, 70.6, 70.5, 49.2, 45.1,37.9, 32.5, 31.4, 29.5, 29.4, 26.7, 25.8, 25.5, 25.0, 22.8, 22.7. MS(ESI); m/z: [M+H]⁺ calcd for C₃₀H₄₃ClN₃O₇: 592.3, found 592.1.

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-6-((5-((6-iodohexyl)oxy)pentyl)oxy)hexanamide

Yield (93%); ¹H NMR (500 MHz, Chloroform-d) δ 9.41 (s, 1H), 8.82 (d,J=8.0 Hz, 1H), 8.30 (s, 1H), 7.71 (t, J=7.8 Hz, 1H), 7.55 (d, J=7.3 Hz,1H), 4.95 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 3.46-3.31 (m, 8H), 3.18 (t,J=6.4 Hz, 2H), 2.94-2.72 (m, 3H), 2.47 (t, J=6.8 Hz, 2H), 2.20-2.12 (m,1H), 1.86-1.72 (m, 4H), 1.66-1.53 (m, 8H), 1.51-1.34 (m, 8H). MS (ESI);m/z: [M+H]⁺ calcd for C₃₀H₄₃IN₃O₇: 684, found 684.0.

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide:DAS-6-5-6-CRBN

Yield (59%); ¹H NMR (500 MHz, CD₃OD/CDCl₃) δ 8.22 (d, J=8.3 Hz, 1H),7.67 (s, 1H), 7.27 (t, J=8.3 Hz, 1H), 7.08 (d, J=7.3 Hz, 1H), 6.86 (d,J=7.3 Hz, 1H), 6.78-6.70 (m, 2H), 5.52 (s, 1H), 4.63 (dd, J=12.0 Hz,J=6.5 Hz, 1H), 3.19 (bs, 4H), 3.00-2.94 (m, 8H), 2.40-2.28 (m, 3H), 2.14(bs, 4H), 2.06-1.98 (m, 7H), 1.86 (s, 3H), 1.74-1.68 (m, 1H), 1.34-1.28(m, 2H), 1.19-1.09 (m, 10H), 1.04-0.85 (m, 8H). ¹³C NMR (151 MHz,CD₃OD/CDCl₃) δ 172.9, 169.7, 168.7, 166.8, 166.1, 162.9, 161.9, 157.1,140.7, 138.9, 137.0, 135.9, 132.8, 131.4, 128.7, 128.1, 126.9, 125.4,125.2, 118.1, 116.2, 82.8, 70.6, 70.52, 70.50, 70.3, 58.3, 52.5, 49.2,43.3, 37.2, 30.9, 29.3, 29.2, 29.0, 27.0, 25.9, 25.8, 25.5, 24.8, 24.6,22.6, 22.4, 17.7. MS (ESI); m/z: [M+H]⁺ calcd for C₅₀H₆₄ClN₁₀O₈S: 999.4,found 999.2.

6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide:BOS-6-5-6-CRBN

Yield (80%); ¹H NMR (500 MHz, CD₃OD/CDCl₃) δ 9.41 (s, 1H), 8.80 (d,J=8.5 Hz, 1H), 8.68 (s, 1H), 7.69 (t, J=8.3 Hz, 1H), 7.51 (d, J=7.3 Hz,1H), 7.48 (s, 1H), 7.43 (s, 1H), 6.92 (s, 1H), 6.88 (s, 1H), 6.49 (s,1H), 4.91 (dd, J=12.0 Hz, J=6.5 Hz, 1H), 4.22 (t, J=6.4 Hz, 2H), 3.77(s, 3H), 3.67 (s, 3H), 3.53-2.46 (m, 14H), 3.41-3.36 (m, 4H), 2.94-2.72(m, 3H), 2.55-2.44 (m, 4H), 2.35-2.32 (m, 1H), 2.17-2.08 (m, 2H),1.79-1.73 (m, 2H), 1.64-1.25 (m, 18H). ¹³C NMR (151 MHz, CDCl₃) δ 168.4,167.3, 165.2, 164.3, 162.8, 150.3, 149.9, 146.3, 145.8, 143.8, 143.5,133.8, 132.9, 132.4, 127.1, 126.6, 121.3, 114.6, 114.4, 113.5, 112.5,111.3, 110.8, 105.7, 101.8, 97.1, 89.9, 68.9, 68.84, 68.81, 66.5, 63.7,54.6, 52.6, 50.8, 49.0, 48.9, 46.8, 45.3, 33.9, 27.5, 25.7, 25.5, 25.4,23.5, 22.6, 22.1, 21.9, 21.0, 18.8, 18.7. MS (ESI); m/z: [M+H]⁺ calcdfor C₅₅H₆₉C₂N₈O₁₀. 1071.4, found 1071.2.

4-((4-(6-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamideIMA-6-5-6-CRBN

Yield (56%); ¹H NMR (500 MHz, CDCl₃) δ 9.40 (s, 1H), 9.24 (s, 1H), 8.81(d, J=8.1 Hz, 1H), 6.70 (dd, J=4.6 Hz, J=1.5 Hz, 1H), 8.55 (s, 1H),8.53-8.49 (m, 2H), 7.97 (s, 1H), 7.84 (d, J=8.1 Hz, 2H), 7.69 (t, J=8.3Hz, 1H), 7.52 (d, J=7.3 Hz, 1H), 7.45-7.40 (m, 3H), 7.32 (dd, J=7.3 Hz,J=1.5 Hz, 1H), 7.21-7.17 (m, 2H), 7.12 (s, 1H), 4.92 (dd, J=12.5 Hz,J=5.4 Hz, 1H), 3.57 (bs, 2H), 3.48 (s, 2H), 3.42-3.36 (m, 8H), 2.91-2.69(m, 4H), 2.52 (bs, 4H), 2.45 (t, J=7.8 Hz, 2H), 2.41-2.35 (m, 2H), 2.34(s, 3H), 2.17-2.12 (m, 1H), 1.76 (quint, J=7.3 Hz, 2H), 1.64-1.43 (m,13H), 1.42-1.27 (m, 6H). ¹³C NMR (151 MHz, CDCl₃) δ 172.2, 171.1, 169.1,168.2, 166.8, 165.4, 162.7, 160.5, 158.9, 151.4, 148.4, 142.0, 137.8,137.7, 136.6, 136.4, 135.0, 133.9, 132.7, 131.1, 130.8, 129.4, 127.0,125.2, 124.4, 123.7, 118.4, 115.4, 115.2, 113.2, 108.3, 70.8, 70.77,70.74, 70.4, 62.4, 58.5, 52.9, 52.8, 50.8, 49.3, 37.9, 31.5, 29.6, 29.5,29.4, 27.4, 26.1, 25.8, 25.0, 22.8, 22.7, 17.7. MS (ESI); m/z: [M+H]⁺calcd for C₅₈H₇₁N₁₀O₈: 1035.5, found 1035.1.

Example

As demonstrated herein, small molecule PROTACs were found to induce thedegradation of BCR-ABL kinase. By varying the target binding ligand(warhead) and the E3 ligase being recruited, BCR-ABL PROTACs can entercells and bind their target, achieving degradation of the protein.

To produce BCR-ABL degrader compounds, BCR-ABL TKIs (imatinib, bosutiniband dasatinib) that bind the c-ABL kinase domain were conjugated to aVon Hippel Lindau (VHL) E3 ubiquitin ligase ligand or to the thalidomidederivative, pomalidomide, to recruit Cereblon (CRBN) E3 ligase. Withoutwishing to be limited by any theory, the resulting bifunctionalcompounds may bind BCR-ABL via the TKI moiety, and bind to VHL or CRBNvia its recruiting ligand. Using the crystal structures of the c-ABLkinase domain in complex with the TKIs (imatinib, dasatinib andbosutinib), positions to attach the linkers to the respective TKIs wereselected such that critical binding interactions were not disrupted(FIG. 1). Four different linkers with varying composition and lengthwere evaluated (FIG. 1C).

These linkers contain a mixture of hydrophobic and hydrophilic moietiesto balance the hydrophobicity/hydrophilicity of the resulting hybridcompounds. The panel of hybrid compounds was then assayed for retentionof binding to c-ABL kinase domain through KinomeScan (Table 1). Allcompounds lost affinity for the phosphorylated and non-phosphorylatedform of ABL compared to the parent compound. Within each warhead series,the 1,5-bis(hexyloxy)pentane linker (hereby designated 6-5-6) yieldedcompounds with the most significant loss (maximum 86-fold) in bindingaffinity (Table 2). All hybrid compounds bound non-phosphorylated c-ABLin the low nanomolar range (0.28 nM-24 nM). The bosutinib- anddasatinib-based PROTACs bound phosphorylated c-ABL in the high picomolarrange (88 μM-1500 μM).

All PROTACs were then tested for c-ABL and BCR-ABL degradation in cellculture. No degradation of BCR-ABL or c-ABL was observed in K562 CMLcells when treated with imatinib-VHL (IMA-VHL) or imatinib-CRBN(IMA-CRBN) PROTACs despite the fact that the PROTACs bound their targetsas evidenced by the reduced phosphorylation of CrkL and STAT5 at higherconcentrations (FIGS. 6A-6B, 7A-7F, 8A-8F, 9A-9F).

A more potent inhibitor warhead (bosutinib or dasatinib) was thenselected for a follow-on PROTAC series. Through a similar syntheticroute, bosutinib was conjugated to the VHL recruiting ligand, producingbosutinib-VHL (BOS-VHL) PROTACs. Despite target engagement as determinedby inhibition of downstream signaling, the BOS-VHL PROTACs also did notinduce degradation of BCR-ABL or c-ABL (FIG. 2A). This finding wasconsistent across several different linkers connecting the bosutinibinhibitor and the VHL recruiting ligand (FIGS. 7A-7F, 8A-8F, 9A-9F). Therepresentative blot shown in FIG. 2A is of BOS-VHL with the linker1-(2-(2-(hexyloxy)ethoxy)ethoxy)hexane hereby designated as 6-2-2-6,which refers to the alkyl/ether composition.

In addition to the BOS-VHL series, dasatinib was also incorporated asthe ligand binding warhead. In contrast to the IMA-VHL and BOS-VHLPROTACs, dasatinib-based PROTAC (DAS-VHL) induced a clear (>65%)decrease of c-ABL at 1 μM PROTAC concentration (FIG. 2B). The apparentdecrease of protein degradation seen at higher PROTAC concentrations (10μM) was observed with other PROTACs and, without wishing to be limitedby any theory, may be attributed to the formation of separatec-ABL-PROTAC and VHL-PROTAC dimers rather than the c-ABL-PROTAC-VHLtrimeric complex required for productive ubiquitination. The c-ABLdegradation seen with the prototype DAS-VHL PROTAC was consistentlyobserved with PROTACs possessing different linkers (FIGS. 7A-7F, 8A-8F,9A-9F). Thus, independent of simple target binding, the inhibitorwarhead (imatinib, bosutinib or dasatinib) largely determines thecapability of a PROTAC to induce c-ABL degradation.

Despite the success of DAS-VHL with the c-ABL degradation, nodegradation of BCR-ABL was seen with any of the VHL-based PROTACs. Thislack of degradation cannot be attributed to loss of binding affinitysince these VHL-based PROTACs still bind and inhibit both c-ABL andBCR-ABL in cell culture (FIGS. 2A-2B). Since E3 ligase presentation tothe target is important for ubiquitination of an available lysineresidue, a differently oriented E3 ligase, such as CRBN E3 ligase, maybe required for adequate ubiquitination and degradation of BCR-ABL.

When dasatinib was conjugated to pomalidomide to recruit CRBN, thedasatinib-CRBN (DAS-CRBN) PROTAC not only retained its ability to inducedegradation of c-ABL (>85% at 1 μM) but also induced BCR-ABL degradation(>60% at 1 μM), demonstrating the first PROTAC-induced degradation of anoncogenic tyrosine kinase (FIGS. 3A-3B).

This result was consistent across the several different linkers usedpreviously in the series of VHL-based PROTACs (FIGS. 6A-6B, 7A-7F,8A-8F, 9A-9F). When the VHL recruiting ligand in the bosutinib PROTACseries was exchanged for the CRBN ligand, c-ABL (>90%) and BCR-ABL(>80%) degradation were observed at 2.5 μM (FIG. 3A). The accessibilityof BCR-ABL and c-ABL for degradation with the BOS-CRBN series stands incontrast with the BOS-VHL series where, despite target engagement, nodegradation of c-ABL or BCR-ABL was observed. Thus, the inactive BOS-VHLcompounds were converted to active BCR-ABL and c-ABL degrader compoundsby switching to the CRBN E3 ligase. As demonstrated by these twoinhibitor warhead series, the oncogenic fusion protein BCR-ABL wasdifferentially susceptible to PROTAC-mediated degradation, depending onthe E3 ligase (VHL or CRBN) recruited to the target.

Since BCR-ABL degradation was observed at 25 nM with theDAS-6-2-2-6-CRBN PROTAC, the cellular effects of the PROTAC wereevaluated (FIG. 3B). In a cell viability assay, DAS-6-2-2-6-CRBN wasactive against BCR-ABL driven K562 with a half-maximal responseconcentration (EC₅₀) of 4.4±2.1 nM (FIG. 4). Furthermore, the PROTACcompound was more than 10³-fold less active against the non-BCR-ABLdriven cell lines, HEK293T and SK-BR-3 breast carcinoma. Thus, thisPROTAC compound retained selective activity against the BCR-ABL drivencell line K562.

TABLE 1 Selected PROTAC Affinities for the ABL Kinase Domain ABL (non-ABL Compound phosphorylated) (phosphorylated) Imatinib 0.86 nM 36 nMIMA-6-2-2-6-VHL 4.3 nM 93 nM IMA-6-2-2-6-CRBN 6.2 nM 110 nM Bosutinib0.063 nM 0.023 nM BOS-6-2-2-6-VHL 1.4 nM 0.63 nM BOS-6-2-2-6-CRBN 0.91nM 0.55 nM Dasatinib 0.03 nM 0.02 nM DAS-6-2-2-6-VHL 0.92 nM 0.47 nMDAS-6-2-2-6-CRBN 0.60 nM 0.32 nM

TABLE 2 ABL (non- ABL Compound phosphorylated) (phosphorylated) Imatinib0.86 nM 36 nM IMA-6-2-2-6-VHL 4.3 nM 93 nM IMA-6-2-2-VHL 5.8 nM 98 nMIMA-6-(2-)₅-2-VHL 1.7 nM 35 nM IMA-6-5-6-VHL 19 nM 46 nMIMA-6-2-2-6-CRBN 6.2 nM 110 nM IMA-6-2-2-CRBN 4.7 nM 72 nMIMA-6-(2-)₅-2-CRBN 3.0 nM 84 nM IMA-6-5-6-CRBN 24 nM 360 nM Bosutinib0.063 nM 0.023 nM BOS-6-2-2-6-VHL 1.4 nM 0.63 nM BOS-6-2-2-VHL 1.3 nM0.67 nM BOS-6-(2-)₅-2-VHL 0.57 nM 0.30 nM BOS-6-5-6-VHL 4.6 nM 1.5 nMBOS-6-2-2-6-CRBN 0.91 nM 0.55 nM BOS-6-2-2-CRBN 0.28 nM 0.11 nMBOS-6-(2-)₅-2-CRBN 0.38 nM 0.13 nM BOS-6-5-6-CRBN 2.8 nM 1.1 nMDasatinib 0.030 nM 0.020 nM DAS-6-2-2-6-VHL 0.92 nM 0.47 nMDAS-6-2-2-VHL 0.36 nM 0.21 nM DAS-6-(2-)₅-2-VHL 0.28 nM 0.088 nMDAS-6-5-6-VHL 2.6 nM 1.3 nM DAS-6-2-2-6-CRBN 0.60 nM 0.32 nMDAS-6-2-2-CRBN 0.28 nM 0.19 nM DAS-6-(2-)₅-2-CRBN 0.26 nM 0.23 nMDAS-6-5-6-CRBN 1.3 nM 1.4 nM

The disclosures of each and every patent, patent application, andpublication cited herein are hereby incorporated herein by reference intheir entirety. While this invention has been disclosed with referenceto specific embodiments, it is apparent that other embodiments andvariations of this invention may be devised by others skilled in the artwithout departing from the true spirit and scope of the invention. Theappended claims are intended to be construed to include all suchembodiments and equivalent variations.

1-24. (canceled)
 25. A method of treating or ameliorating a tyrosinekinase-dependent cancer in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of at least one compound of formula (I):TKI-L-(ULM)_(k)  (I), wherein: TKI is a tyrosine kinase inhibitor, L isa linker, each ULM is independently a ubiquitin ligase binder, and k isan integer ranging from 1 to 4, wherein TKI is covalently linked to Land wherein each ULM is covalently linked to L; or a salt, enantiomer,stereoisomer, polymorph or N-oxide thereof.
 26. The method of claim 25,wherein the cancer is associated with overexpression and/or uncontrolledactivation of the tyrosine kinase.
 27. The method of claim 25, whereinthe tyrosine kinase is oncogenic.
 28. The method of claim 25, whereinthe subject is a human.
 29. The method of claim 25, wherein the cancercomprises chronic myelogenous leukemia (CML).
 30. The method of claim25, wherein the compound is administered to the subject by at least oneroute selected from the group consisting of nasal, inhalational,topical, oral, buccal, rectal, pleural, peritoneal, vaginal,intramuscular, subcutaneous, transdermal, epidural, intrathecal andintravenous routes.
 31. The compound of claim 25, wherein the TKI iscapable of binding to at least one of c-ABL and BCR-ABL.
 32. The methodof claim 25, wherein the tyrosine kinase-dependent cancer is associatedwith overexpression or uncontrolled activation of at least one of c-Abland BCR-ABL, wherein the TKI is capable of binding to at least one ofc-ABL and BCR-ABL.
 33. The method of claim 32, wherein the tyrosinekinase-dependent cancer is c-ABL-dependent cancer or BCR-ABL-dependentcancer.
 34. The method of claim 33, wherein the cancer comprises chronicmyelogenous leukemia (CML).
 35. The method of claim 32, wherein thecompound is administered to the subject by at least one route selectedfrom the group consisting of nasal, inhalational, topical, oral, buccal,rectal, pleural, peritoneal, vaginal, intramuscular, subcutaneous,transdermal, epidural, intrathecal and intravenous routes.
 36. Themethod of claim 25, wherein the TKI is Imatinib, Dasatinib, orBosutinib.
 37. The method of claim 25, wherein at least one ULMcomprises formula (IX):


38. The method of claim 25, wherein at least one ULM comprises formula(X):


39. The method of claim 25, wherein k is
 1. 40. The method of claim 25,wherein the compound is selected from the group consisting of:N-(2-chloro-6-methylphenyl)-2-((6-(4-((S)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-11,14,17-trioxa-4-azatricosan-23-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-6-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-22-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-2-2-6-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-VHL):

(2S,4R)-1-((S)-3,3-dimethyl-2-(2-(2-((6-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-2-2-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-((S)-3-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carbonyl)-2,2-dimethyl-5-oxo-7,10,13,16,19,22-hexaoxa-4-azaoctacosan-28-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-2-2-2-2-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-27-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-2-2-2-2-2-2-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-((5-((6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-5-6-VHL):

(2S,4R)-1-((S)-3,3-dimethyl-2-(6-((5-((6-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(IMA-6-5-6-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-22-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-4-oxo-10,13,16-trioxa-3-azadocosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-2-2-6-VHL):

(2S,4R)-1-((S)-2-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-2-2-VHL):

(2S,4R)-1-((S)-2-(tert-butyl)-27-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-azaheptacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-2-2-2-2-2-2-VHL):

(2S,4R)-1-((S)-2-(6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)hexanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide(BOS-6-5-6-VHL):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)ethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-6-CRBN):

6-(2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide(BOS-6-2-2-6-CRBN):

4-((4-(6-(2-(2-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)ethoxy)ethoxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide(IMA-6-2-2-6-CRBN):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-CRBN):

2-(2-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide(BOS-6-2-2-CRBN):

4-((4-(6-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-2-oxoethoxy)ethoxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide(IMA-6-2-2-CRBN):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(1-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-1-oxo-3,6,9,12,15,18-hexaoxatetracosan-24-yl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-2-2-2-2-2-2-CRBN):

24-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-3,6,9,12,15,18-hexaoxatetracosanamide(BOS-6-2-2-2-2-2-2-CRBN):

N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-24-(4-(4-((4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)carbamoyl)benzyl)piperazin-1-yl)-3,6,9,12,15,18-hexaoxatetracosanamide(IMA-6-2-2-2-2-2-2-CRBN):

N-(2-chloro-6-methylphenyl)-2-((6-(4-(6-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide(DAS-6-5-6-CRBN):

6-((5-((6-(4-(3-((3-cyano-4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxyquinolin-7-yl)oxy)propyl)piperazin-1-yl)hexyl)oxy)pentyl)oxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)hexanamide(BOS-6-5-6-CRBN):

4-((4-(6-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)-6-oxohexyl)oxy)pentyl)oxy)hexyl)piperazin-1-yl)methyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)benzamide(IMA-6-5-6-CRBN):


41. The method of claim 25, wherein the compound is administered to thesubject as a pharmaceutical composition further comprising at least onepharmaceutically acceptable carrier.
 42. The method of claim 25, whereinthe subject is further administered at least one additional therapeuticcompound that treats or ameliorates the cancer.